Difference in reproductivity and offspring growth between litter numbers in four strains of mice

As a means to improve mouse production, the differences among litter numbers in reproductivity, post-weaned growth and puberty of offspring were examined in 4 strains of mice, SHN/Mei, SLN/Mei, C3H/HeMei and GRS/AMei. Females were mated to be concurrently pregnant and lactating and reproductivity was recorded from the 1st to the 3rd litters. Some offspring of both sexes were weighed at 30, 40 and 50 days of age and vaginal opening was also checked in females. Pre-weaned growth of offspring differed little between litter numbers in all strains except GRS/A. Meanwhile, post-weaned growth of offspring was higher in the 2nd and 3rd litters than in the 1st litter of SHN and SLN, associated with a decreased rate of still-born pups. In C3H/He, post-weaned growth of offspring was higher in the 1st and 2nd litters and rearing rate decreased for the 3rd litter. In GRS/A, the 1st litter showed the greatest pre- and post-weaning growth rate of offspring. All findings indicate that in SHN and SLN strains the later litters are better for offspring production, while in C3H/He and GRS/A strains it is the earlier litters. The results stress the value of having information on reproductive characteristics of mouse strains for efficient offspring production.     

Mammary tumour incidence in relation to the pattern of oestrous cycles in mice

The patterns of oestrous cycles were examined in new strains of mice exhibiting a high (SHN) and a low (SLN) mammary tumour incidence. These mice originated from the same stock of Swiss albino mice. At the age of 3-4 months, SHN mice showed much longer cycles with continual dioestrous periods than SLN mice. The average lengths of cycles and dioestrous periods were, respectively, 8.3 +/- 0.3 and 6.0 +/- 0.4 days in SHN mice and 4.6 +/- 0.1 and 2.5 +/- 0.1 days in SLN mice. Furthermore, the distinct alteration in the patterns of cycles from SHN to SLN type was associated with complete reduction in mammary tumour incidence in highly inbred C3H/He female mice maintained by brother X sister mating with no selection for mammary tumourigenesis. The relation between the pattern of oestrous cycles and mammary tumourigenesis is discussed from the viewpoint of the endogenous hormonal milieu and the responsiveness of target organs.     

Relationship between milk composition and pup's growth in mice

The effect of lactation numbers (first, second, and third) on milk composition and the relationship between milk composition and pup's growth were examined in four strains of mice (SHN, SLN, C3H/He, and GR/A). Although the percentage of milk fat and moisture increased and decreased, respectively, through the first to third lactations in C3H/He mice, no differences in milk composition among lactation numbers were observed in the other mouse strains. In pooled data derived from the four strains of mice, there was no significant correlation between milk composition and body weight or growth rate of pups on Day 12 or 20 of the first lactation. In contrast, the amount of milk fat and moisture was the positively and negatively, respectively, correlated with the pup's growth parameters during the second and third lactations. This was reflected by a lower fat content and a higher moisture content in the milk of GR/A mice, especially evident during the second and third lactations and reduced pup's growth, when compared with the other mouse strains. Milk lactose and ash content was not correlated with pup's growth parameters. The results of our study stress the importance of milk fat for the growth of mouse pups.     

Effects of concurrent pregnancy and lactation on reproduction in four strains of mice

The influence of concurrent pregnancy until the third lactation on reproduction was studied in four strains of mice (SHN, SLN, C3H/He and GRS/A) as a possible step to evaluate the efficiency of this system on offspring production. In all strains, little difference was observed between sequential and concurrent pregnancy and lactation groups in any parameter as the index of reproductivity--delivery interval, litter size, average weight and growth rate of pups, mother weight, rearing rate, still-birth rate and rate of still-born pups. Also, no difference was seen between groups of any strain, in the number of females which had three litters. The estimated number of pups weaned per female during 50 days after first being placed with males was higher in the concurrent pregnancy groups than in the sequential pregnancy groups of all strains. Meanwhile, the incidence of spontaneous mammary tumors, the major characteristic of these strains, was consistently lower in the concurrent pregnancy group in all strains. These results have demonstrated that the concurrent pregnancy and lactation system is an efficient means of mouse production.     

Horizontal transmission of the mouse mammary tumor virus in cage mates of the same and opposite sex of low and high mammary cancer strain mice

Horizontal transmission of mouse mammary tumor virus (MTV) was investigated in cage mates of the same and opposite sex of low (BALB/c) and high mammary cancer strains (DD/Tbr, SHN and GR). By MTVp27 and MTVgp52 radioimmunoassay, MTV antigen expression was found in the salivary glands, mammary glands and secondary male genital organs of the MTV-free BALB/c strain. Infectivity and oncogenicity were also found in DDf or BALB/c mice by inoculating extracts of salivary gland and/or seminal vesicle in high mammary cancer strains. It is suggested that the primary source of the infectious agent in cases of caged animals of the same sex is saliva, while the primary source in cases of caged animals of the opposite sex is the seminal fluid, although additional infection through saliva cannot be ruled out in the latter case.     

Effect of chronic suppression of growth hormone secretion, by SMS 201-995 on lactation in mice

The effect of chronic suppression of growth hormone (GH) secretion by SMS 201-995 on lactation was studied in primiparous C3H/He mice. Mammary gland DNA content on day 12 of lactation was significantly lower in SMS 201-995 treated mice than in the control. There were little differences between groups in mammary gland RNA content and litter growth on day 12 of lactation. That was associated with a slightly higher RNA/DNA ratio and a significant increase in food intake during lactation. These results indicate that inhibited mammary gland growth by GH suppression has little effect on lactation. The smaller mammary gland can compensate by increasing its secretory activity.     

Mammary gland growth and response to hormones in mastomys compared with mice

Mammary gland growth with or without hormone manipulation was examined in virgin mastomys (Praomys (mastomys) natalensis) and compared with C3H/He mice having a low mammary tumor incidence. Mammary glands of mastomys consisted mostly of duct systems even at 720 days of age, whereas conspicuous formation of normal end-buds and preneoplastic hyperplastic alveolar nodules were seen in the glands of mice after 180 days of age. Mammary glands of mastomys showed a higher response to estrogen or progesterone rather than prolactin, which is much different from other rodents.     

Effects of Ovariectomy and Prolactin on Mammary Gland Expressions of Transforming Growth Factor alpha and Epidermal Growth Factor Receptor mRNAs in Mice

Beginning 15 days after ovariectomy (OVX), a high mammary tumor strain of SHN virgin mice at 3 months of age received subcutaneous injections of danazol (0.5 mug / 0.1 ml olive oil, once a day), perphenazine (0.05 mg / 0.1 ml saline, twice a day) or ovine prolactin (oPRL: 0.25 mg / 0.05 ml buffer, twice a day) for 3 days to modulate their circulating PRL levels. The serum PRL level was significantly decreased by danazol and increased by perphenazine compared to the intact and OVX-control groups. The expression of both transforming growth factor alpha (TGFalpha) mRNA and epidermal growth factor receptor (EGFR) mRNA in the mammary gland was increased by danazol. However, TGFalpha mRNA expression was decreased by perphenazine. Meanwhile, mammary end-bud formation was inhibited in danazol-treated group. All findings suggest that the manifestation of the effect of TGFalpha on mammary gland is rather suppressed by PRL, while mammary gland growth needs the participation of PRL; in other words, PRL is dominant to TGFalpha on the mammary gland growth. OVX resulted in a significant decrease of TGFalpha mRNA expression in the mammary gland despite of little alteration in serum PRL, confirming the previous observations. The similar trend was observed in ICR mice; however, the response to hormonal modulation is generally less susceptible than SHN mice.     

A device for milk collection from mice

Approximately 14 days after SHN, SLN and C3H/He strains of mice gave birth, their pups were removed. After 14--16 hours away from their pups, the dam was anesthetized, injected with oxytocin and milked. The milking device consisted of a rubber-stoppered centrifuge tube containing a stainless steel catheter and polyvinyl chloride tube. It was operated by vacuum.     

Two inbred strains of mice with high and low mammary tumor potentials established from the same basal stock of Swiss albino (SHN and SLN): history of mammary tumorigenesis and reproduction after the 30th generation of full-sib mating.

We established SHN and SLN strains of mice with a high and a low mammary tumor potentials from the same basal stock of Swiss albino. The selection history of females (mothers) for subsequent generations, mammary tumorigenesis and reproduction up to recent generations are presented and compared with the results up to F30 published in 1979. The changes with generation of each parameter varied in both strains except the conditions of female (mother) selection for subsequent generations, which were generally constant. However, the fluctuations in parameters up to F30 became generally smaller with advancing generations. Based on the fluctuations from generation to generation of mammary tumorigenic and reproductive parameters, an interdependency between mammary tumor potential and reproductivity is suggested in SLN, but less so in SHN. The pup growth rate was increased and associated with selection for mammary tumorigenesis in SHN; this parameter is usually used as an indicator of mother lactational performance in mice. The significance of selection for mammary tumor potential in association with reproductivity was further discussed.     

Incomplete proviral genome of mouse mammary tumour virus is present on chromosome Y of NIH Swiss and some genetically related mouse strains.

An incomplete proviral genome of endogenous mammary tumour virus (MMTV) was found in DNA of several strains of mice. This MMTV-related sequence was assigned to the Y chromosome since it was clearly observed in male mice only. This MMTV provirus contained a sequence related to LTR (long terminal repeat), but not to gag-pol and env genes. NFS, NIH Swiss/S, STS/A, and DD/Tbr mice have this sequence but BALB/cHeA, SHN, SLN, C57BL/6NJcl, C3H/HeNJcl and CBA/JJcl mice are negative. In the strains containing this sequence, a DNA test for the sequence makes it possible to easily distinguish the DNAs of male or female mice.     

Genetic marker patterns and endogenous mammary tumor virus genes in inbred mouse strains of Japan

In order to establish the genetic relatedness of the inbred mouse strains kept in Nara, genetic marker patterns were determined in conjunction with a study on endogenous mammary tumor viral genes in these strains. Isoenzyme patterns combined with patterns of other genetic markers, show that the unrelatedness between various inbred strains of the dd stock is as high or even higher as between strains of known different origin and geneology. Based on endogenous viral gene patterns the dd stock derived mice can be subdivided into three group, DDD, DDN, DDO, KF and DD/Tbr. The DD/Tbr and its foster-nursed substrain (DD/Tbrf) have the lowest number of endogenous viral genes, i.e. two, while the other strains carry 4-6 such genes. The SLN and SHN strains, derived from a Swiss stock, have a similar pattern of viral genes different that of all other strains studied, also strains of Swiss origin from other sources, such as the NFS and the GR.     

Enhancement of spontaneous mammary tumorigenesis at advanced age in mice by temporary stimulation of mammary growth during youth

As a possible step to clarify the relationship between mammary gland growth during youth and mammary tumorigenesis at advanced age, the long-term effects of the temporary stimulation of mammary gland growth at youth on spontaneous mammary tumorigenesis was studied in a high mammary tumor strain of SHN virgin mice. They received a single pituitary graft under the kidney capsule or a subcutaneous implantation of progesterone for 60 days between 30 and 90 days of age. Both treatments enhanced mammary gland growth and the effect of pituitary grafting was higher. Spontaneous mammary tumorigenesis paralleled mammary gland conditions at youth. The results indicate that the stimulation of mammary gland growth during youth enhances spontaneous mammary tumorigenesis at advanced age in mice, which is quite contrary in rats, and they explain the cause of the higher mammary tumor potential in breeders than in virgin mice.     

A novel mechanism of resistance to mouse mammary tumor virus infection

Exogenous mouse mammary tumor virus (MMTV) is carried from the gut of suckling pups to the mammary glands by lymphocytes and induces mammary gland tumors. MMTV-induced tumor incidence in inbred mice of different strains ranges from 0 to as high as 100%. For example, mice of the C3H/HeN strain are highly susceptible, whereas mice of the I/LnJ strain are highly resistant. Of the different factors that together determine the susceptibility of mice to development of MMTV-induced mammary tumors, genetic elements play a major role, although very few genes that determine a susceptibility-resistance phenotype have been identified so far. Our data indicate that MMTV fails to infect mammary glands in I/LnJ mice foster nursed on viremic C3H/HeN females, even though the I/LnJ mammary tissue is not refractory to MMTV infection. Lymphocytes from fostered I/LnJ mice contained integrated MMTV proviruses and shed virus but failed to establish infection in the mammary glands of susceptible syngeneic (I x C3H.JK)F(1) females. Based on the susceptible-resistant phenotype distribution in N(2) females, both MMTV mammary gland infection and mammary gland tumor development in I/LnJ mice are controlled by a single locus.     

Coexpression of exogenous and endogenous mouse mammary tumor virus RNA in vivo results in viral recombination and broadens the virus host range.

Mouse mammary tumor virus is a replication-competent B-type murine retrovirus responsible for mammary gland tumorigenesis in some strains of laboratory mice. Mouse mammary tumor virus is transmitted horizontally through the milk (exogenous or milk-borne virus) to susceptible offspring or vertically through the germ line (endogenous provirus). Exogenously acquired and some endogenous mouse mammary tumor viruses are expressed at high levels in lactating mammary glands. We show here that there is packaging of the endogenous Mtv-1 virus, which is expressed at high levels in the lactating mammary glands of C3H/HeN mice, by the virions of exogenous C3H mouse mammary tumor virus [MMTV(C3H)]. The mammary tumors induced in C3H/HeN mice infected with exogenous MMTV (C3H) virus contained integrated copies of recombinant virus containing a region of the env gene from an endogenous virus. This finding indicates that there was copackaging of the Mtv-1 and MMTV(C3H) RNAs in the same virions. Moreover, because Mtv-1 encodes a superantigen protein with a V beta specificity different from that encoded by the exogenous virus, the packaging of Mtv-1 results in an infectious virus with a broader host range than MMTV(C3H).     

Binding of glucocorticoid receptors to mammary chromatin acceptor sites

We have recently characterized the interaction of mouse mammary estrogen receptors (ER) with mammary chromatin acceptor sites and demonstrated that ER from estrogen resistant lactating mammary glands do not bind to chromatin. In this study we have characterized the chromatin binding of the glucocorticoid receptor from mouse mammary glands isolated from nulliparous and lactating mice in order to better understand the relationship between receptor binding to chromatin and steroidogenic sensitivity of the tissue. Mammary chromatin was linked covalently to cellulose and deproteinized sequentially by 0-8 M Gdn-HCl. Binding to intact chromatin as well as to chromatin deproteinized by Gdn-HCl was determined using partially purified [3H]dexamethasone labelled glucocorticoid-receptor complexes (GR) obtained by fractionation on DEAE-cellulose columns. The binding of [3H]GR from mammary glands of nulliparous mice to chromatin fractions from the same tissue revealed maximal binding activity (acceptor sites) on chromatin previously extracted with 5-6 M Gdn-HCl. Binding of [3H]GR was of high affinity (Kd = 0.2 nM) and saturable. A simultaneous comparison of the chromatin binding patterns for [3H]ER and [3H]GR isolated from mammary glands of nulliparous mice revealed that the chromatin subfractions obtained with 4-6 M Gdn-HCl extraction contained acceptor sites for both [3H]ER and [3H]GR; however, while the [3H]ER bound to a 4.5 M and a 5.5 M site, the [3]GR bound a 5 M and a 6 M site. Competition experiments supported the steroid receptor specificity of the chromatin acceptor sites. Thus, the 4-6 M chromatin fractions contain distinct acceptor sites for the glucocorticoid receptor and for the estrogen receptor. In addition our studies reveal that the binding patterns of [3H]GR isolated from mammary glands of nulliparous and lactating mice to their homologous chromatin is essentially similar. Thus, in contrast to estrogen receptors, glucocorticoid receptors from lactating mammary glands are able to effectively bind to chromatin acceptor sites which supports our previous suggestion that the estrogenic insensitivity of lactating mouse mammary glands may at least be in part due to the impeded interaction of ER with chromatin acceptor sites.     

Increased sensitivity of mouse mammary gland for hormones and tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate in tissues containing the Mtv-2 gene

We have compared the response of mammary explants obtained from GR mice with those derived from GR/Mtv-2- congenic mice towards hormones and tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). In explants of the mammary glands of virgin GR mice, insulin, prolactin, progesterone and TPA stimulated the rate of (3H)-thymidine incorporation into DNA. A significantly lower response towards prolactin and TPA was observed in mammary explants derived from GR/Mtv-2-mice. The differential sensitivity of mammary tissues from GR and GR/Mtv-2-mouse strains to the actions of prolactin and tumor promoter is directly correlated to their different incidences of spontaneous tumor formation; it is not known if there is a causal relationship.     

Pituitary prolactin and growth hormone levels during different reproductive states in mice with a high or a low lactational performance

The pituitary prolactin and growth hormone (GH) levels were determined by disc electrophoresis on 10% polyacrylamide gel during the virginal and pregnant stages and on Day 12 of lactation, using C3H/He and C57BL/6 mice. The former had been shown to be superior to the latter in both mammary development and lactational performance. The pituitary prolactin levels were significantly higher in C3H/He mice than in C57BL/6 mice during the virginal and pregnant stages. However, no strain differences existed in the prolactin levels on Day 12 of lactation. Little difference in the prolactin levels was found between estrus and diestrus, and the levels declined gradually with the advance of pregnancy in both strains. The levels decreased after 1 hr of suckling preceded by 8-hr removal of young on Day 12 of lactation in both strains, but the difference between before and after suckling was statistically significant only in C3H/He mice. Both pituitary GH content and concentration were significantly higher in C3H/He mice than in C57BL/6 mice during the virginal stage and the content was also higher in C3H/He mice during the pregnant stage. However, there existed no strain difference in the levels on Day 12 of lactation. Little change in the pituitary GH levels was observed during the different reproductive states in both strains.     

Development of a cellular immune response in a highly cancer-susceptible C3H/He strain of mice and its nonfactor substrain C3H/f after the transplantation of a syngeneic mammary gland tumor]

Syngeneic mammary gland tumor (MMT-1) grew slower in non-factor C3H/f (MTV-L) mice than in wirus-positive C3H/He (MTV-S) mice. The immunization of mice with tumor (MMT-1), inoculated and later excised, revealed a higher immune reactivity in the mice of the non-factor substrain. In studying the dynamics of the cytotoxic activity of lymphocytes in regional lymph nodes cytotoxically active lymphocytes were found to appear in C3H/f mice on day 6 after the inoculation of tumor (MMT-1), while the areactive state of lymphocytes was observed in C3H/He mice on days 3 and 24 and in C3H/f mice on day 24 after the inoculation of tumor (MMT-1). Spleen lymphocytes in C3H/f mice had no cytotoxic effect on tumor cells.