Maternal Inheritance and my Mitotyping Project

My interest in maternal inheritance has been longstanding. My
mitochondrial DNA study project has been contemplated for over
fifteen years before I got brave enough to think I might be able to
make some kind of positive contribution in knowledge toward the
Thoroughbred that I dearly love. It was Charles Bruce Lowe whose
writings and difficulties gave me inspiration and confidence to at
least make the attempt. An abortive attempt was made in 1997 with
Dr. Ann Bowling, but she suddenly passed away at the age of 41 from a
cerebral hemorrhage.

I have prepared myself by operating a Thoroughbred breeding farm for
36 years, immersing myself in pedigree theories, conducting breeding
experiments on my farm, breeding research, biological research, and a
background of extensive historical research. My planned matings
using pedigree patterns, inbreeding, balanced-sex inbreeding,
techniques learned from successful breeders, and voracious reading of
Hewitt, Lowe, etc. has produced very satisfying and successful
runners in a regional setting.

Our mtDNA study was started by, financed by, and matured with a horse
breeder interested in selective mating, maternal inheritance, and the
expression of performance traits. I have tried to NOT bring to the
table the baggage of stubborn opinion or "know-it-all" closed
mindedness. I hope I have the open mindedness that objective science
requires. Academic pursuit of knowledge while important is parallel
to the potential benefit of pragmatic application of modern,
biological science for the horse breeder's benefit. In this genomic
age we are still applying eighteenth century, archaic breeding
practices.

Mitochondrial investigation must be more than a tool of identity in
order to be incorporated by the interested horse breeder in his
breeding program. I am happy and willing to acknowledge the great,
groundbreaking contribution made by Hill, Cunningham, Bowling, et al.
I have no intent to duplicate their efforts - why would I want to do
what has already been done?

Our interest in identifying matrilines is just an initial step
because how can legitimate research be done if you cannot identify
that which you are studying? How can you trust any conclusions if
you don't know from where you started? Obviously, therefore, the
research must start with identity. In approximately 20 -25 percent
of the samples we have sequenced so far, the mtDNA haplotype is in
disagreement with the recorded lineage of the individual. The Jockey
Club had no parentage verification prior to 1987 other than an honor
system dependent on the integrity of Thoroughbred breeders.
Registration relied on written and diagrammatic descriptions of
external markings similar to those used by state and federal live
stock inspectors with other domesticated animals. From 1987 until
2001, parentage verification was determined through nuclear DNA from
a sample of blood in addition to the written and diagrammatic
descriptions (except for an unsatisfactory attempt to use nasal swabs
for DNA verification in 1994-95). The integrity of lineages is
crucial to any rational selective breeding scheme. Mis-
identification of Thoroughbred lineages has lent an unwelcome and
unpredictable variability to selective mating schemes causing
inexplicable and unexpected failures from some matings selected by
experienced and expert breeders. Mis-identification of lineages
invalidates any thoughtful, objective attempt at breeding selection.

Our ultimate objectives are not the same as previous researchers. We
have better equipment [Transgenomics WAVE technology], a state-of-the-
art, forensic genetics laboratory, at least 8x better accuracy with
our methods compared to previous methods, far cheaper and quicker
processing, etc. At this time, we are building our own DNA database.
Access to the hair and blood samples retained by The Jockey Club
would be a researcher's dream.

In Mendelian inheritance, the nuclear DNA is the scaffolding or
structure. There are little differences at the level of DNA between
a horse, a human, or a hamster. What goes on is going on beneath.
Researching the nuclear genome is so large and complex that it would
take millions of dollars and far more elaborate facilities, years,
and resources to make any kind of meaningful contribution - Inquiry
into nuclear inheritance is far beyond my limited means.

The much smaller mitochondrial genome of "Equus caballus" [ACCESSION
X79547] is 16660 bp [base pairs] in size. It is monoclonal [that is
it replicates or clones from dam to progeny. The mitochondrial
genome does not split, divide, and recombine as in nuclear
processes]. It is somewhat simpler to understand and research.
PLUS, it has to do with maternal inheritance, which is where my
interests have laid for thirty years. Long live the ladies!
Yours truly, Loren

Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 farm
505-604-2221 cel
lbolinger1@comcast.net
-or-
lbolinger@gmail.com

Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Mitotyping Project

Mitotyping Project
By Loren Bolinger 9/14/2005

Unlike previous scientific inquiries, my first impulse was to present
our project directly to the typical Thoroughbred breeders across the
USA and internationally. The Internet and several breeders' e-mail
discussion groups proved useful to query a wide range of breeders
directly at minimum cost. I was curious as to the interest of other
Thoroughbred breeders to my passion and obsession. I also thought
that it would be more likely to pick up a wider range of unusual
matrilines, scientifically interesting, and suitable candidates for
sampling. Some of the matrilines are rare or obscure and more likely
to be found in the hinterlands. These rare matrilines may have
important relevance to our project.

One, very satisfying observation I've made, is the surprising numbers
of horse breeders sophisticated about maternal lineages,
mitochondrial inheritance, the Lowe family numbers of their horses,
and the potential of my mitotyping project to not only accurately
resolve known identity problems, discover unknown, longstanding mis-
identifications, but also for its application by horse breeders to
future selective matings. All horse breeders passionately want to
improve their stock. Accurate identification of lineage is crucial
to many selective breeding schemes.

The response to my mtDNA inquiry has been warmly rewarding to me
through the very large volume of e-mail and overall enthusiasm of
horse breeders internationally. I had planned to place small
advertisements in industry publications to get a wider range of
samples, but now must wait additional funding.

I have received tremendous grassroots response to what some might
regard as a rather obscure, technical inquiry. I can assure the
Jockey Club that my informal and anecdotal survey reveals that a
majority of Thoroughbred breeders are, in fact, tremendously
interested in the potential of mitochondrial research, more accurate
identification of maternal lineages and therefore, more accurate
identification of most all Thoroughbreds in the current population to
the current limits of modern scientific methods. The biological
knowledge and advances in scientific technologies are accelerating at
a mind-spinning rate - new discoveries coming almost day-by-day. We
are in a new golden age of genomic discovery. It would be unfair to
the breed we love to not take advantage of these advances to preserve
and conserve the Thoroughbred horse. For the most part, we still use
archaic selective breeding practices at a time when the biological
sciences are making major contributions the health and welfare of
humans and many species of domesticated and wild plants and animals.
If the knowledge learned from mitotyping could be used to reduce the
needless wastage and injury, minimize genetic defects and diseases,
conserve the breed for the future, while improving selective mating
techniques that produce the modern Thoroughbred racehorse, the effort
and expense would be well worth it. To my mind, it is a win - win
situation.

I strongly believe that ultimate responsibility for the
identification of the Thoroughbred lies with the keepers of the
General Stud Book, such as the Jockey Club and other international
keepers of the studbooks in their countries. They have the existing
recorded data, the authority, and the resources. There are an unknown
number of undiscovered haplotypes in lineages of the current
Thoroughbred population. One of my intentions is to attempt to
discover as many of the unknown haplotypes as my means allow; the
purpose being to make known the entire number of surviving haplotypes
comprising the modern Thoroughbred breed. I believe this is an
important project and I hope that my tiny mtDNA study will inspire
and encourage the appropriate authorities to create and fund an
international project to identify and verify all Thoroughbreds to the
limits of our technologies as far back as possible.

Yours truly,
Loren

Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Thoroughbred horses as unique research subjects

Thoroughbred horses as unique research subjects:

An Ideal Model for Studies of Inheritance and Selective Mating

By Loren Bolinger

I have been a Thoroughbred horse breeder for over 36 years in New
Mexico. I have a passion for selective mating, historical research,
and improvement of the breed. I believe that improvement to the
breed can be made by breeders through selective mating. Many horse
breeders since the founding of the breed have had an anecdotal belief
in maternal inheritance. In 1894, Charles Bruce Lowe classified and
ranked Thoroughbreds by matrilineal lineage and each family's success
at elite races. His was also the first widely publicized [among
Thoroughbred horse breeders], systemic attempt to document and prove
maternal inheritance and the validity of certain forms of selective
mating. During the 1960's biologists discovered the importance of
the mitochondrial genome and established incontrovertible proof of
the second system of biological inheritance - maternal inheritance
through DNA located in the cytoplasm of the ovum. Mitotyping is the
term used for methods that identify the true matriline of the
individual under examination. Errors in Thoroughbred parentage
classification of many historical individuals' lineages have
propagated through entire branches of their family trees to the
present GSB, and even when known, have never been possible to verify
and corrected.

Now that there are accurate, reproducible, and cost effective
techniques, it is especially important to breeders to verify and
correct matrilineal lineages. Any sort of reasonable attempt at
breed improvement through selective mating depends on accurate
lineages.
Goals include reconnection of the lost American, Australian Colonial,
Argentinean families with their English roots [and other unidentified
members of the current Thoroughbred breeding population]. Another
goal is to untangle, separate, and correct the misidentified,
incorrectly recorded English lineages that occurred primarily in the
1800's. Yet another goal is to assess the connection between
mitochondrial inheritance and the expression of performance traits.

Dr. Phillip W. Danielson, University of Denver and Dustin Mark
Gilbert, our research assistant, are skilled and highly trained in
DNA forensics. Dr. Danielson conducts training seminars for law
enforcement and investigative personnel in crime scene procedures.
Their protocols and procedures are accurate, rigorous, comprehensive,
and reproducible. The mitotyping we use produces cutting edge
accuracy and reproducibility [better than 99.9 per cent accuracy].
They have trained me in mtDNA collection, data collection, and
fieldwork. Our DNA technology is far cheaper, faster, and more
accurate. We are using the same procedures used in criminal
forensics. Our lab is currently processing about 35 - 50 samples per
week with only one research assistant.

1. Our laboratory procedures used for processing buccal swabs, blood,
and hair samples incorporates duplicate methods used by the US Jockey
Club for parentage verification and registration. Our hope is to
eventually gain access to the DNA repository of the Jockey Club to
continue this important research.

2. Our procedures and protocols are compatible with those used by Dr.
Emmeline Hill reported in Animal Genetics, August 2002 [pp. 287-294].
We have verified and reproduced results as reported in "History and
Integrity of Thoroughbred Dam Lines Revealed in Equine mtDNA
Variation."

Why the Thoroughbred
The Thoroughbred horse represents an unusual research opportunity
with potentially far reaching implications for all species. Nearly
all plants and animals chosen for research are relatively randomly
mated, wild mated, mated without human-caused selection, or mated
with selection for simple or single gene traits. Often,
inappropriate "selfing" or other radical selection practices are used
in the laboratory and often on strains of animal models completely
divorced from the real world.

In contrast, among all domesticated animals, the Thoroughbred is
mankind's longest lived and most successful, selective-mating
experiment. It has had a relatively consistent goal over
approximately three hundred fifty to four hundred years - the test of
the winning post. That the selective mating strategies have been
successful can be assessed by the physical changes in the organism
since its founder stock. The phenotypes of the modern day
Thoroughbred, evolved over about 350 - 400 years, can also be
compared against those of other breeds of horses. This particular
breed, developed through specialized selective mating, performance
testing, and culling, is unique among all domesticated animals.

It is generally agreed that multi-factorial genes (and possibly)
maternal inheritance control traits involved with elite performance.
Research into inheritance of multi-factorial traits has been
extremely difficult and complex. Traits inherited through the
mitochondrial genome are not well understood. The Thoroughbred
brings a unique genetic background that is better suited to the
unraveling of the conundrum of understanding complex traits. It is
unique in the organization, selection, and culling of its lineages
and individuals. Historically, all forms of selective mating
experiments have been tried.

The large body of Thoroughbred horse breeders is highly motivated to
improve horse breeding techniques yet for the most part biological
research has ignored the Thoroughbred. The horse among domesticated
animals has had a far stronger economic impact today than the
proportionate research dollars spent on its behalf would indicate.
The North American Thoroughbred industry has an economic impact as
reported in the Wednesday, June 29, 2005 electronic edition of the
Thoroughbred Times of $39.2 billion , an economic impact study in
Australia revealed the contribution of the Australian Horse Industry
was estimated at 6.3 billion in 2001 , and in Great Britain, in 1999
[most recent year], it was estimated that the economic impact of the
British horse industry was about 2.5 billion, second largest economic
activity in the British countryside only to farming , yet
Thoroughbred breeders have received little attention from the
biological sciences. We are basically still using crude, eighteenth-
century selection practices and/or many breeders are governed by non-
genetic issues such as commercial appeal, fad, and fissionability.
[The element of uncertainty added by misclassification of lineages
should be unacceptable to horse breeders intent on breeding the very
best individuals possible incorporating selective-mating methods with
correction, verification, and finally accuracy of lineages using
scientific objectivity from the genomic era. An unacceptable
variable can be easily eliminate by modern biological science.]
Current Progress
Dr. Emmeline Hill [et al 2002] and Dr. Phillip Danielson [Gilbert,
Bolinger et al 2005 preliminary] have shown that mitochondrial DNA
analysis is an easy, cost effective, accurate, and reproducible way
to identify and characterize horses by their families or maternal
lineages.

Analysis by both groups [above] has shown large discrepancies in the
GSB [General Stud Book] classifications. Most errors seem to have
arisen from misclassification of horses very early in the bloodlines
of the Thoroughbred.

There seems to be far fewer founding mares [matriarchs] than
originally believed due to convergence of many families into one
haplotype and the fact that diverging families seem to result from
misclassification rather than multiple founding mares or mutations.

Lastly, the founding mare population was more than likely small due
to the very limited and controlled breeding by the early English
founder breeders in the initial stages of the species' development.

With our limited, current database, our first goal is to continually
gather more data, especially on families that have not yet been
sequenced.

The next step while we continue to enlarge our database would be to
begin tracking down American, Australian, Argentinean, and other lost
bloodlines and connecting them back to their English ancestors.

Extremely valuable horses are being created with 19th century
selection methods. Far too much wastage and injuries occur, and
needless, catastrophic breakdowns occur. In the amazing explosion of
the current Genomic Era, it seems relevant to employ biological
discoveries in breed improvement, conservation, humane treatment, and
preservation of the Thoroughbred. The possibilities exist that
significant progress can be made to reduce unsoundness and wastage in
the Thoroughbred through breakthroughs in scientific selective
mating.

We wish to understand the contribution toward elite athletic
performance that comes through maternal inheritance - the
mitochondrial DNA contained in the cytoplasm that is inherited only
through the tail-female lineage. In order to accomplish this
objective, the accuracy of the lineages must be first restored or
corrected. Until this is done, selective mating schemes are useless.
If I didn't have the help of a top genetic laboratory and one of the
best forensic geneticists in the United States, Dr. Phillip
Danielson, this project would never have been possible.

My personal aim is to disclose the pure research, hopefully, if it
has any merit at all, published in a scientific journal and on the
TBHeritage website, like the other matriline discoveries. The goal is
to be able to produce practical results that breeders can apply, not
ivory tower research that only gathers dust and cobwebs and only
satisfies intellectual curiosity. I'm a pragmatist, I wholeheartedly
believe in maternal inheritance with respect to performance traits,
have put my limited resources where my mouth is, and incorporated my
beliefs in my breeding program. I am satisfied [at my level of
participation in the horse breeding business] the results I am
getting. It would be very satisfying to have biological evidence to
back up my passion for breeding horses.

Yours truly,
Loren
Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Accumulated inbreeding over several generations

Accumulated inbreeding over several generations.doc By Loren
Bolinger, 10/17/2001

1

Accumulated inbreeding over several
generations

A tail-female bloodline, or matriline, composed of a series of
relatively-inbred broodmares from a successful matriline that
has reliably exhibited high levels of blacktype performance
(even if success is somewhat distant) has been performance-
sensitized. That is, genetic conditions have been constructed
(or set in place) that channel the forces of inheritance toward
continued winning performance from descendants from this
matriline. Mares descended from such a family will often
continue successful productivity of performance traits even
when bred to regional sires that are relative outcrosses,
provided that there is, at least, limited inbreeding within six
generations to a few key ancestors. Limited, near inbreeding
to prepotency is more effective when the distant strains in the
sire and dam are organized. Distant kinship is one of the
forces of inheritance successfully used by Bruce Lowe, both in
Australia for Frank Reynolds and others and with his
nineteenth century importations for James Ben Ali Haggins’
Rancho del Paso Stud in California.

Selection and mating of breeding stock related through distant
kinship is a great organizational tool that results in increased
expression of performance traits. As in other mating schemes,
selecting stallions that have already produced high quality
progeny and/or have exhibited the highest possible racing class
will increase the probabilities of producing successful progeny.
Mating breeding stock whose own phenotypes tend toward the
mean phenotype for their particular bloodline will reproduce
their bloodlines’ characteristics more reliably. Breeding
extremes to one another tends to be extremely unreliable and
genetically more variable. Stallions and mares should have
compatible phenotypes.

In planning matings for future progeny from relatively inbred
female families, the breeder should key off certain prepotent
selected ancestors in the pedigree of the broodmare. Even
with limited inbreeding to only a few key ancestors, mostly
outcrossed progeny from inbred broodmares can still express
performance traits. In other words, a small boost from
relatively limited reinforcement of only a few ancestors will
cause the re-expression of performance in these progeny
beyond the mean for the breed. Sire-candidates selected for
such broodmares even though near outcrosses, should have
distant kinship to the tail-female family of the broodmare.
While using six-generation hypothetical breedings (on paper or
the computer screen) to determine the feasibility of the mating,
extended 8 –10 generations or more will assist in assessing
distant relationships of the proposed mating.

It is more desirable, if at least two of the broodmares in the
first four or five dams of the tail-female series have relatively
balanced inbreeding within six generations. The accumulations
of relatively balanced inbreeding should occur in at least
several of the broodmares in the series (usually closer to the
target, better in the dam and the granddam). The breeder
should prefer reinforcements of duplicated ancestors within
six-generations that have built-up or accumulated in at least a
minimum of two dams.

Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Fixing The Type

1

FIXING THE TYPE
by Loren Bolinger, 2/27/04 - 3/3/04

How the early breeders fixed the type of the
Thoroughbred: Intermingled colateral matrilines, and
melded early Founder patrilines. How their
stewardship of the breed was so successful that today's
horsemen are still reaping the rewards of their
pioneering selective mating techniques. The science of
genetics has acknowledged and proved the validity of
many of the early techniques.

The breeder manipulated the genotype through selective
mating techniques, typically inbreeding and the
recognition and repetition of certain pedigree patterns.
The breeder selected and culled though evaluation of
the resulting offspring of the mating primarily through
phenotype measured by objective performance and/or
its relationship to performance.
Fixing the Type (establishing the typology)
The Thoroughbred horse is a hybrid breed that normally does
not breed true-to-type. The racehorse breeder attempts to
reduce the phenotypic volatility or variability in the racehorse
he breeds through employing mating strategies that increase
multiple instances of inbreeding or linebreeding over
successive generations, while culling out those individuals that
fail to meet expectations. Inbreeding reduces the number of
ancestors in a pedigree by duplicating those thought to be
more important or prepotent. By purifying the ancestry, the
breeder reduces the variability, hopefully to increase the
probability of breeding true-to-type.]

Fixing the type means to reinforce a family’s or strain’s
phenotype through selective breeding, heavy overall
inbreeding, and culling. The purpose of fixing the type is to
increase the permanence and stability of the genotype so that
the breeder can more reliably predict the phenotype and to
more reliably trigger the expression of performance traits in the
offspring resulting from selective matings. By increasing
stability of the genotype through careful reduction of the total
number of ancestors, variability of the phenotype can be
dramatically reduced. Selective breeding techniques have been
devised that attempt to more reliably predict and reproduce
desired inherited traits and characteristics in offspring from
selective matings without significantly increasing deleterious
possible side effects of inbreeding. Typology means the
phenotypic or physical characteristics that are manifested from
an offspring’s genotype and the environment with which it
interacts.
How Fixing the Type was Acomplished

Selective mating was accomplished through three general
strategies: Genetic Isolation - Differentiation [breeding toward
a desired goal] cannot take place unless no new genetic
material is introduced over a period of time. No additional
founder-type bloodlines were introduced, once the original
male and female Founder stock was chosen. Artificial
Selection - The breeder selects matings between individuals
exhibiting desired traits or attributes and avoids random
matings. Inbreeding - Inbreeding reduces diversity [weeds out
undesirable traits] while fixing or reinforcing desired traits.
In order to “fix type,” a breeder or several generations of
breeders must closely linebreed among the descendants of
several, selected, prepotent individuals that are complementary
to each other and in whom a high rate of successful offspring
are produced. In this way, the breeder concentrates the
successful and productive genetic assets of a family. The
breeder selects patterns that sex-balance the immediate
descendants of duplicated ancestors, selects mates in such a
way that patterns of inversion in their bloodlines-in-common
occurs along paternal and maternal wings, inbreed to great
mares and to great matrilines. The breeder selects prepotent
stallions from compatible bloodlines, and avoids, as much as
possible, excess male inbreeding while simultaneously
increasing inbreeding to females, and increases the overall
quality and quantity of matriarchs/great mares in the pedigrees
of the offspring. The above are some of the selective mating
strategies that lead to success. The final strategy, sometimes
emotionally difficult, is to cull for failure. Cull for failure to
perform, cull for deviation from phenotype. Selective mating
means to select mates based on the most reasonable
hypotheses and objectives concerning bloodlines, pedigree
patterns, and phenotype. Those that fail to live up to the
objectives should be culled from the selective breeding
program.
Pedigree Matching
Bloodlines colateral to each other share an important role in
the selective mating technique called pedigree matching, that
is, utilizing those bloodlines that developed in parallel with one
another. Pedigree matching means that ancestors of the
colateral bloodlines share kinship or relatedness with one
another through powerful ancestors-in-common. The hooks of
the key members of the colateral bloodlines were commonly
opposite-sex-balanced - ancestral links or hooks inverted by
sex from each other.

Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Inbreeding vs Diversity

Inbreeding vs Diversity
Argument [7/10/2005]
While inbreeding generally results in a decrease in diversity, I
believe this statement must be qualified. Male inbreeding,
especially excess male inbreeding over several generations results
in a decrease of diversity and it also tends to adversely affect the
ability to express or trigger the expression of performance traits, it
may well also increase a tendency for conformational flaws.

Balanced-sex inbreeding has a much more moderate effect on
diversity and does not diminish it in the same way as excess male
inbreeding. Inbreeding to female influences somewhat inexplicably
strengthens existing diversity possibly through increased
preservation [reinforcement] of matrilines. The effect may be due
to redressing the [ratio] balance between the unequaled
reproductive productivity between males (breeding many) and
females (producing few) in favor of the female (through the
reinforcement of maternal inheritance).

-Loren Bolinger
Loren Bolinger
Running Horse Farm LLC
7102 Ilfield Road Southwest
Albuquerque, New Mexico 87105-7046
505-873-2220 Voice
505-877-4443 FAX
e-mail: lbolinger1@comcast.net

Loren Bolinger, boss TBMitotyper

This blog was created to support my Thoroughbred
mitochondrial DNA project. The mtDNA study was
contemplated for twelve or fifteen years before I got
brave enough to think I might be able to make some
kind of positive contribution in knowledge toward the
Thoroughbred horses that I dearly love.

It was Charles Bruce Lowe in 1895 whose writings
and difficulties gave me inspiration and confidence to
at least make the attempt. I have prepared myself by
operating a Thoroughbred breeding farm for 34 years,
immersing myself in pedigree theories, conducting
breeding experiments on my farm, breeding research,
biological research, and a background of extensive
historical research.

Our mtDNA study was started by, financed by, and
matured with a horse breeder interested in selective mating,
maternal inheritance, and the expression of performance
traits. I have tried to NOT bring to the table any
baggage of stubborn opinion or "know-it-all" closed
mindedness. I hope I have the open mindedness that
objective science requires. Academic pursuit of knowledge
while important is parallel to the potential benefit of
pragmatic application of modern, biological science for
the horse breeder's benefit.

Mitochondrial investigation must be more than a tool of
identity in order to be incorporated by the interested
horse breeder in his breeding program. I am happy and
willing to acknowledge the great, ground-breaking
contribution made by Hill, Cunningham, Bowling, et al.
I have no intent to duplicate their efforts - why would
I want to do what has already been done?

Our interest in identifying matrilines is just an initial
step because how can legitimate research be done if you
cannot identify that which you are studying? How can you
trust any conclusions if you don't know from where you
started? Obviously, therefore, the research must start
with identity.

In Mendelian inheritance the DNA is the scaffolding or structure.
There are little differences at the level of DNA between a horse,
a human, or a hamster. What goes on is going on beneath.

Researching the nuclear genome is so large and complex
that it would take millions of dollars and far more
elaborate facilities, years, and resources to make any kind of
meaningful contribution - Far beyond my limited means.

The much smaller mitochondrial genome of "Equus caballus"
[ACCESSION X79547] is 16660 bp [base pairs] in size. It is
monoclonal [that is it replicates or clones from dam to progeny.
The mitochondrial genome does not split, divide, and recombine
as in nuclear processes]. It is somewhat simpler to understand
and research. PLUS, it has to do with maternal inheritance
which is where my interests have laid for thirty years. Long
live the ladies!!
-Loren Bolinger