Chemical evidence of the quantitative thiamine and riboflavin requirements of the rat during pregnancy and the development of the fetus

162 Pages·013.859 MB·English

by  Barrett, Margaret Louise

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6BBXZ0AL EVIDENCE
OF fill QUANTIfAflfl THUMBS AMD 1IB0FMVII SBOKHfflBMBHTS
OF TBS SAT SUKXSO PREGNANCY AMD TUI
XBEVBLOPMSHT OF TBS FETUS
Margaret liaise Barrett
A Dissertation Submitted to the
{graduate Faculty in Partial Fulfillment of
the Requirements for the Degree of
DOCTOR m PBXLOSQFBT
Major Subject! iutrltion
Approved!
In Gharge'bf Major Work
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Heal r©F Sa jo^Wpar^en|T'
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Dean1" of lradu£te Coliege "
Iowa State College
1950
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UMI Number: DP12357
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TABLE Of CONTENTS
Pag®
INTRODUCTION................. 1
PART X . . . . . . . . . . . . . . . . . . . . . . . . 6
EMBRYOLOGIC AL BlflLOPJIlNf OF f 11 R A T ............ 6
Ovulation, Maturation, and Fertilization , , 9
Pronuolear to Blastocyst Stag© ........... 11
Implantation to th® Tenth B a y . 18
Tenth Bay {1 to 19 Somites)........... * * 30
Eleventh Bay (20 to 26 Somites) . . . . . . 38
Twelfth Bay (27 to 33 Smites)......... . 40
Thirteenth Bay (34 to 47 Somites) . . . . . 42
Fourteenth Bay (48 to 80 Somites) . . . . . 43
fifteenth Bay (61 to 85 Somites) . . . . . . 44
Sixteenth Day 45
Seventeenth D a y ................ 43
Eighteenth Day . . . . . . . . . . . . . . . 48
nineteenth Bay to Tew . . . . . . . . . . . 46
The Uterus 47
EXPERIMENTAL PROCEDURE . . . . . . . . . . . . . 49
Animals and Their Daily Care . . . . . . . . 50
Breeding Technique and Establishment of
the Time of Initiation of Pregnancy . . . 50
Experimental Sroupa . . . . . . . . . . . . 51
The Ration .............. . . . . . . . . . 52
Preparation of Tissues for Future
Vitamin Assays . . . . . . . . . . . . . . 53
RESULTS AND DISCUSSION ............ . . . . . . 56
SUMMARY AND G OUGHTS IGJIS . . . . . . . . . . . . . 80
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Pag#
PART II 81
fJHAMIlB RBQDIKBIISRV8 DURING HmCBOdVXGV ..... 82
' SXFSRXMQnVAL FROCBDBR* . . . . ......... . . 86
thiamine Assay .Procedure . . . . . . . . . . . 86
The Metabolism Studi@a . . . . . . . . . . . 87
RlitfLTS AMD DISCUSSIO* . ........ ........... f 1
■Deposition of Tfal&ala# la Tissues at
a Mean# of Determining Thiamine
Requirements for'Reproduction.......... 91
■■ Metabolism.Experiment® Conducted on
Pregnant and ion-pregnant tats as
a leans of Determining Thiamin©
Requirements for Reproduction . . . . . . 101
SUMMARX AID CCBCLUSIOHS .......... 117
PARf 111 . . . . . . . . . . . . . . . . . . . . . . * 120
RIBOFLAVIN BBQPX RUSKS DOR1IO REPRODUCTION . . . 121
EXPERIMENTAL P1001DTOE . . . . . . . . . . . . . . 126
tibofl&vin Assay Procedure . . . . . . . . . 126
RESULT® AID Di®6U»l« 127
'Deposition ©f Riboflavin in Maternal
and Petal tissues as a leans of
Determining Riboflavin Requirements
for Reproduetloa 127
Metabolism Experiments Conducted on
Pregnant and Ion-pregnant lats as
a Means of Determining' Riboflavin
Requirement® for Reproduction . . . . . . 154
Sfnous* AID OOtfGEOftXm . . . . . . . . . . . . . . 147
HTIRATDil CITED . . . . . . . . . . . . . . . . . . . . 14®
AOXIORlBDCnims * . . . . . . . . . . . . . . . . . . . 166
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iMfmmcnm
Numerous studies In the field of nutrition have
suggested a ©lose relationship- he tween the maternal diet and
the sondition of the ehiid at birth. Faulty diet prior to
and during pregnancy Is believed to be responsible for sev­
eral types of disorder® encountered at this time. Miscarri­
ages, prematurity, malformations of the young, stillbirths,
and toxemias of pregnancy have been noted more frequently
among women whose diets were judged Inadequate in various
essential nutrients .than among women with excellent dietary
Intakes» fhese statements are born# ©at by the findings of
Ebbs and his associates (1941), Burke, Beal, Kirkwood and
Stuart (1943), and Balfour (1944).
It is encouraging that the number of deaths of the
mother and Infant caused by disorders of pregnancy has been
steadily decreasing in the past decade, though this is true,
vital statistics for the United States for the year 1947
revealed that there were 23.7 stillbirths per 1000 live
births, that prematurity was the most serious cause of infant
mortality, and that congenital malformations and congenital
debility of unknown origin were frequently observed (U. S.
Public le&lth Service, 1949). fhe infant mortality rate for
the United States as a whole was 32.2 per 1000 live births,
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the lowest rate la Met©ry,. fills average figure represents
a rat# of 50*1 for the white population* 47.7 for negroes*
and 65.7 for non-whit® Infants including the American Indian,
Chin#®#* and. Japanese*
While the stodies by 'Burk# and her associates as well as
those of many others have been ©specially valuable sine© they
have been conducted on human subjects, they are only a begin­
ning to a complete understanding of the importance of good
nutrition during pregnancy, f© date little Is known about
the specific factors which prevent the various disorders.
Likewise,' we have very little information ©oncoming the
quantitative needs of nutrients at different stages of
pregnancy.
Animal .experimentation* conducted under more, rigidly
controlled conditions has also stimulated a keen interest In
the importance of good nutrition during pregnancy and in the
possibility that certain of the B-vitasiiaa may be important
in the prevention of abnormalities which are occasionally
observed In infanta. The extensive studies of Warkamy and
his co-workers (194B, 1943* 1944) have shown that riboflavin
is a vital-factor in the normal development of the embryo*
When inadequate amounts of this vitamin were consumed by
the mother, malformations of the palate, syndactylism, club­
feet, and shortening of certain bones- became apparent in
the young. Recently two other publications have suggested
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that m&lform&tlons of the newborn are dm to deprivation of
certain B-vltamlns* Boisselot (1948) has observed defective
fetuses as a result of to© little pantothenic acid in the
maternal diet* O’Dell and Hogan (1950) have reported a high
incidence of hydrocephalics among young rats bom to animals
fed a folic acid inhibitor* Withdrawal of biotin (Kennedy
and Palmer, 1945), pantothenic acid (MeIson and Ivans, 1946),
and B^g (Baerson et al„, 1949 j Bear, I960) likewise have been
found to induce disorders of pregnancy.
■ With .these challenging observations In mind, plans'were
mad® for a rather extensive investigation of the importance
of the B**vitaain® during pregnancy. While these experiments
were to be Halted t© reproduction in the albino rat for the
present, it was hoped that any special stress periods
observed for the rat would be considered critically for human
reproduction*. Iventually deficiency studies of the B-vitamins
were anticipated to determine whether inadequate amounts of
these factors might bring to light additional malformations
in the offspring which might aid in explaining defects
observed in Infants. Such information should ultimately lead
to their prevention*
As. a. beginning to this larger program, it was believed
advisable to study the normal stock animal during pregnancy
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and to observe both the rat® of prenatal development of the
young and the rate of deposition of certain vitamins in
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plaosntal and fatal tissues at Intervals during' the gesta­
tion- period, this Information should refleet the vitamin
needs ©f the mother-during pregasacy and should make'us aware
of fluctuations la these needs at different intervals of
pregnancy.
the' present study has heea divided into thro®' major
sections* 'Fart 1 include®' a 'detailed review of the liters- •
ture describing the' embryonic development of the rat* Data
are presented on the prenatal growth of normally developing
young of stook animals produced in the Sutrition Laboratory
of the'Food® and nutrition department* Fart XI of the study
Includes Information on the occurrence of thiamine in fetal
tissues and of the thiamine store* of the female throughout
pregnancy* A limited'number of observations are also
presented on the' urinary excretion of thiamin# of pregnant
and non-pregnant ilttermates* 'these findings have been
included since they offer an additional mean* of estimating
the thiamine requirement during reproduction. Part III of
this .Investigation deals with the deposition of riboflavin
in maternal and fetal tissues* This section likewise
include® data on the excretion of riboflavin by the kidney.
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