Normal mice develop from oocytes injected with spermatozoa with grossly misshapen heads.

Because in the mouse some structurally abnormal spermatozoa can penetrate oocytes, we decided to determine whether mouse oocytes fertilized by spermatozoa with grossly misshapen heads are able to develop normally. We injected such spermatozoa from BALB/c mice individually into mature oocytes of hybrid mice B6D2F1. Eighty to ninety percent of the oocytes were activated and developed into blastocysts in vitro and, when transferred to foster mothers, some developed into apparently normal fertile adults. This finding indicates that a proportion of abnormal spermatozoa carry all the genome and organelles necessary for normal embryonic development and growth to fertile maturity.

Effect of culture systems on mouse early embryo development.

The amount of oxygen available to embryos in drop cultures under paraffin may be limited by the gas mixture employed, the method of equilibration or by the use of positive or negative pressure filtration. When these factors were varied, in experiments using Swiss outbred (SO) embryos, the only significant difference detected was the poorer development of embryos from the 1-cell to blastocyst stage when cultured in medium without prior equilibration. Also under these conditions, the inclusion of glucose in the medium did not increase the incidence of 2-cell block, and glutamine in the presence of glucose enhanced the development of 1-cell embryos to blastocysts. One-cell (C57BL/6 x SJL) F1 x SO embryos were successfully cultured in HEPES buffered CZB medium with added bicarbonate, under an atmosphere of air. The proportion of blastocysts formed in this medium, with a concentration of 10 mM sodium bicarbonate, was not different from that developed in CZB under 5% CO2 in air.

Sex reversing non-disjunction of the Y chromosome produces exceptionally low sex ratio (% males) and hermaphrodites in the progeny of male BALB/cBm mice: the roles of the maternal genotype and the Y chromosome.

When females of 21 strains and hybrids were mated to BALB/cBm males to determine the role of the maternal genome in the sex reversing non-disjunction of the Y chromosome, (1) BALB/cBm and BALB/cBy and SJL/J females produced 39.5-41.5% males and 2.4-2.8% hermaphrodites; (2) SWR/J, A/HeJ, DBA/2J and C3HeB/FeJ produced 44.8-49.1% males and 0.2-0.7% hermaphrodites; (3) C3H/HeJ and three strains of C57BL produced normal sex ratios and no hermaphrodites; (4) four F1 hybrids produced 44.5-49.2% males and 0.3-1.9% hermaphrodites; (5) the seven CXB RI strains produced perplexing sets of data: 26.5%-52.0% males and 0.2-3.2% hermaphrodites. These results indicate that a partly dominant gene favouring non-disjunction occurs in the female genomes of BALB/c and SJL/J strains, an enhancing gene occurs in C57BL/6By and there may be others. Heterosis appears to favour normal mitosis. CXBH females produced 26.5% males and 3.2% hermaphrodites, indicating that non-disjunction may have occurred in every male zygote, thus providing models for the generation of Turner's syndrome, hermaphroditism and a predictable non-disjunction. Reciprocal crosses were made between SJL/J and BALB/cBm, followed by 20 backcrosses to the maternal strains, to exchange the Ys and produce two new consomic strains. Males from SJL-BALB/cBm-Y strain, when mated to CXBH females, sired 34.3% males and 4.3% hermaphrodites, whereas BALB/cBm-SJL-Y sired no hermaphrodites and the sex ratio of the offspring was normal. This shows that the non-disjunction involves only the BALB/cBm Y chromosome and is completely independent of genes on the X or autosomal chromosomes. These results indicate that the BALB/cBm Y chromosome is unable to interact normally with the mitotic spindles of some genotypes, particularly CXBH, BALB/c and SJL. The simplest hypothesis is that a primary non-disjunction occurs at first cleavage. This can produce an array of mosaics determined by chance in the many sampling events that take place during development and by the relative vigour and stability of the two original clones.

Do histocompatibility genes influence sex ratio (% males)?

Breeding records for 15 of the 42 C57BL/10SnJ Congenic Histocompatibility Mouse Strains from the Special Mouse Stocks Resource (SMSR) showed three with a significant excess of male offspring. Strain B10.R111(71NS), hereafter R, gave the highest proportion of males (55.68 +/- 0.59% of 7129 newborn) which is significantly more than the proportion of male offspring (49.81 +/- 0.94% of 2853 newborn) from the C57BL/10SnJ progenitor strain, hereafter B. All mice in the SMSR colonies were fed Old Guildford 96 and Old Guildford 96W liberally on alternate weeks. Breeding females of B and R strains were established in a research colony at the Jackson Laboratory under conditions similar to those in SMSR except that they were fed Wayne Sterilizable Rodent Blox. More than 5000 inbred, hybrid and backcross fetuses were examined but no evidence of an excess of males was found. Also, there were no strain differences in the neonatal data. However, the sex ratio of the 4396 neonates was just significantly higher (P less than 0.05) than that of the fetuses, indicating some perinatal loss of females. An even greater loss of females was most probably the cause of the high sex ratios in the preliminary and follow-up surveys of SMSR R mice, which we ascribe to an interaction between the H-2 haplotype of the R strain, or a gene linked thereto, and the Old Guildford diet that is unfavourable to female survival. The sex ratio of fetuses agreed so closely with the Mendelian expectation as to indicate that the primary sex ratio was 50% males and that the R strain is not a model for the human male bias. There were no hermaphrodites or twins and the sex ratio of the fetuses in litters without resorptions was normal. These findings emphasize the variability of presumptive secondary sex ratios.

Spontaneous malignant granulosa cell tumors in ovaries of young SWR mice.

Granulosa cell tumors (GCT) of the ovary appear spontaneously at 4-6 weeks of age in SWR/J and in SWR/Bm inbred strain mice, with a maximum incidence reached by 10 weeks. Cancer was confirmed by metastasis to abdominal organs and by transplantability of primary tumors to histocompatible hosts. Results of genetic crosses showed that GCT appear in SWR X SJL F1 but not in SJL X SWR F1 nor in other F1 females derived from matings of SWR mice with A/HeJ, C57BL/6By, CBA/J, or DBA/2J mice. These findings suggest the maternal transmission of GCT susceptibility. Recombinant inbred strains SWXJ were produced from a progenitor mating of a SWR female to a SJL male. At F20, females in 3 of 14 SWXJ strains developed GCT, with one strain displaying a 5-fold increase in incidence. Embryo transfer studies with SWXJ-6 and -9 mice suggested that maternal transmission was most likely via the fertilized egg rather than through milk or placenta-uterine contact. Analysis of metaphase chromosomes indicated that the modal number in tumors and bone marrow was 40 (2n = 40) with 2 X chromosomes present. Gross chromosomal aberrations were not detected. A working hypothesis proposes that interaction of a unique SWR factor, perhaps cytoplasmic, with nuclear genomic material common to Swiss mouse stocks results in occurrence of GCT in young SWR and SWR-derived mice.

A cytogenetic investigation of inherited true hermaphroditism in BALB/cWt mice.

A predictably high incidence of true hermaphroditism (3%) among fetal BALB/cWt mice provided the opportunity for studying the cause of inherited hermaphroditism. G-banded liver metaphase chromosomes were analyzed from 20 fetal hermaphrodites and from 15 normal female and male littermate controls. All hermaphrodites and seven males were chromosomal mosaics, i.e. XO/XY or XO/XY/XYY. In each mosaic fetus there were more XO than XYY cells. From these findings we hypothesize that BALB/cWt hermaphrodites arise from non-disjunction and probable loss of the Y-chromosome during mitosis. Results from breeding experiments indicated that a defect in the BALB/cWt Y-chromosome per se accounts for its nondisjunction and loss. A significant relationship was found between the amount of ovarian tissue and the percentage of XO cells in mosaic fetuses. If all chromosomally mosaic (XO/XY and XO/XY/XYY) individuals are considered, ovarian tissue was formed if there was 24% or more XO cells in an individual. If only hermaphroditic individuals are considered, ovarian tissue was formed if there were 44% or more XO cells. Finally, we found that the distribution of ovarian and testicular tissue within an ovotestis of BALB/cWt hermaphrodites was not random: ovarian tissue was situated at both ends or only at the cranial end; in only one case was ovarian tissue situated in the center flanked by testicular tissue. We suggest that such nonrandom distribution of ovarian and testicular tissue argues against the hypothesis that the organization of testicular tissue is dependent only on the presence of H-Y antigen.

The morphology of fetal gonads of spontaneous mouse hermaphrodites.

The gonads of 107 spontaneous, chromosomal mosaic, day-15 fetal hermaphrodites derived from BALB/cWt strain male mice are described and photographs of seven gonads representative of the major types are presented. There were 16 ovaries, 15 testes, and 183 ovotestes. The ovotestes contained on the average more testicular tissue than ovarian, and the ovarian tissue was more frequently located at the gonad poles, particularly the craniad pole. There was no difference between left and right sides with regard to gonad type, but more pure gonads were found on the left than on the right side (21/10), Meioses were observed throughout the ovarian tissue and also in some testicular cords, particularly in the caudad pole of the gonad. Some meiotic figures contained sex vesicles. The significance of these findings is discussed in relation to several aspects of sex determination.

Chemical Scent Constituents in the Urine of the Red Fox (Vulpes vulpes L.) During the Winter Season.

Four volatile chemical compounds have been identified as apparently unique constituents in urines of red foxes (both sexes) during the winter season when mating occurs. Quinaldine was found only in male fox urine. Several other compounds identified are found in other species also. Some or all of these compounds may function in olfactory communication in the red fox.

Identification of dihydrothiazoles in urine of male mice.

The compounds 2-isopropyl-4,5-dihydrothiazole and 2-sec-butyl-4,5-dihydrothiazole were identified in urine of male mice. Their excretion in mouse urine is sex-dependent. In the urine of female mice the two dihydrothiazoles are either absent or present only in trace amounts. The analytical procedure includes the adsorption of the volatile mouse urine constituents on Tenax GC, their gas chromatographic separation and the detection of the sulfur compounds by the sulfur-specific detector. The identified compounds were synthesized. Natural and synthetic compounds had the same mass spectrometric properties and retention data.