Description

The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.

Strain Information

Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Species laboratory mouse   Donating Investigator Deborah Good,   Virginia Polytechnic Institute

Description
Male mice homozygous for this targeted mutation exhibit bilateral cryptochism, hypogonadism, azoospermia, low testosterone and follicle stimulating hormone levels and lack instinctual male sexual behavior. Homozygous females are hypogondal unless raised in the presence of male mice, have estrous cycle defects, and reduced fertility. No gene product (mRNA) is detected by Northern blot analysis of embryos. Initially, homozygous male mice, 4 to 7 weeks of age, have a reduced weight compared to wildtype controls. By 12 weeks of age, homozygous males are significantly heavier than wildtype, and mutants progressively increase weight with age. Female homozygotes 5 to 6 weeks of age have normal body weight, however by 7 weeks of age they are significantly heavier than wildtype controls. The increased body weight is due to increased adipose tissue, with the exception of female homozygotes more than 52 weeks of age that exhibit increased lean body mass. Male heterozygotes and homozygotes, between 12 and 19 weeks of age, are polyphagic. Female homozygotes, older than 52 weeks of age, have significantly lower body temperature than wildtype. Male homozygotes between 5 and 10 weeks of age and female homozygotes between 7 and 11 weeks of age exhibit reduced voluntary physical activity. Heterozygotes are viable, fertile and male heterozygotes display adult onset obesity. This mutant mouse strain may be useful in studies of the endocrine hypothalamic-pituitary axis, fertility, onset of puberty, adult-onset obesity and metabolism.

Development
A targeting vector containing neomycin resistance and herpes simplex virus thymidine kinase genes was used to disrupt the entire coding region. The construct was electroporated into 129S4/SvJae derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice.

Control Information

	Control
	Wild-type from the colony
	101043 B6129SF1/J	(approximate)
Considerations for Choosing Controls

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms provided by MGI

      assigned by genotype

Nhlh2^tm1Irk/Nhlh2⁺

        involves: 129S4/SvJae * C57BL/6

• growth/size phenotype
• increased susceptibility to age related obesity
  • male heterozygotes weigh significantly more than wild-type males at 12-25 weeks of age, with the exception of week 19; by 52 weeks, male heterozygotes are significantly heavier than wild-type males (average 46.52 g vs 39.07 g, respectively) with significantly more body fat   (MGI Ref ID J:96351)
  • unlike males, female heterozygotes show normal body weights at 11 and 25 weeks of age, and are not significantly heavier than wild-type females at 52 weeks   (MGI Ref ID J:96351)
• adipose tissue phenotype
• increased percent body fat
  • at >52 weeks, male (but not female) heterozygotes show a significantly increased percent body fat relative to wild-type littermates, indicating a sex difference   (MGI Ref ID J:96351)
• behavior/neurological phenotype
• abnormal motor capabilities/coordination/movement
  • unlike male heterozygotes, female heterozygotes are comparable to female homozygotes with respect to daily voluntary activity, showing a significant reduction of daily wheel rotations/stage relative to wild-type females   (MGI Ref ID J:96351)
• polyphagia
  • at 12-19 weeks of age, male (but not female) heterozygotes exhibit significantly increased food intake over wild-type males   (MGI Ref ID J:96351)

Nhlh2^tm1Irk/Nhlh2^tm1Irk

        involves: 129S4/SvJae * C57BL/6

• mortality/aging
• early reproductive senescence
  • most mutant females exhibit premature reproductive senescence and support only 3-4 pregnancies during the first few months of a 9-month period   (MGI Ref ID J:101925)
• behavior/neurological phenotype
• abnormal mating receptivity
  • both young and aged female homozygotes show at least a 50% reduction in hormone-stimulated sexual behavior as measured by their lordosis quotient   (MGI Ref ID J:101925)
• abnormal voluntary movement
  • at 5-10 weeks of age, preobese male homozygotes display significantly reduced voluntary running wheel activity relative to wild-type males   (MGI Ref ID J:96351)
  • similarly, at 7 and 11 weeks of age, female homozygotes exhibit significantly reduced voluntary running wheel activity, regardless of the stage of the estrous cycle   (MGI Ref ID J:96351)
  • however, no differences in circadian rhythm, balance, or learning of motor tasks are observed   (MGI Ref ID J:96351)
• polyphagia
  • at 12-19 weeks of age, male (but not female) homozygotes exhibit significantly increased food intake over wild-type males   (MGI Ref ID J:96351)
• reduced male mating frequency
  • male mice did not copulate   (MGI Ref ID J:39356)
• digestive/alimentary phenotype
• abnormal anogenital distance
  • slight reduction in the distance from the penis to the anus   (MGI Ref ID J:39356)
• endocrine/exocrine gland phenotype
• *normal* endocrine/exocrine gland phenotype
  • serum levels of growth hormone (GH) and luteinizing hormone (LH) were normal in male mice   (MGI Ref ID J:39356)
• • abnormal Leydig cell morphology
    • reduced number of leydig cells at 32 weeks of age   (MGI Ref ID J:39356)
  • abnormal seminal vesicle morphology
    • convoluted   (MGI Ref ID J:39356)
  • absent corpus luteum   (MGI Ref ID J:39356)
  • cryptorchism
    • bilateral cryptochism   (MGI Ref ID J:39356)
  • impaired ovarian folliculogenesis
    • arrested at the early stages of antral formation   (MGI Ref ID J:39356)
  • seminiferous tubule degeneration
    • only remnants of the tubules remained at 32 weeks of age   (MGI Ref ID J:39356)
  • small male preputial glands
    • preputial glands were either small or absent   (MGI Ref ID J:39356)
  • small ovary
    • females raised in the absence of males exhibited small and pale ovaries   (MGI Ref ID J:39356)
    • size reduction was not apparent until after 4.5 weeks of age   (MGI Ref ID J:39356)
  • small seminiferous tubules
    • rudimentary lumens   (MGI Ref ID J:39356)
  • small testis
    • significant reduction after 8 weeks of age, though apparent at 4.5 weeks of age   (MGI Ref ID J:39356)
    • testicular tissue was largely replaced by a capsule of fat at 32 weeks of age   (MGI Ref ID J:39356)
• growth/size phenotype
• increased susceptibility to age related obesity
  • at 4-7 weeks of age, male homozygotes show a significant weight reduction relative to wild-type males; however, starting at 10-12 weeks, male homozygotes show a progressive increase in body weight that is equivalent to 14% and 33% over wild-type at the age of 25 and 52 weeks, respectively   (MGI Ref ID J:96351)
  • unlike males, female homozygotes exhibit normal body weights at young ages (5-6 weeks), but become significantly heavier than wild-type females by 7 weeks of age   (MGI Ref ID J:96351)
  • with the exception of female homozygotes which show increased lean body mass at >52 weeks of age, increased body weight is due to increased body fat, and not due to overall increases in lean mass, body size, or length   (MGI Ref ID J:96351)
  • adult-onset obesity is preceded by reduced voluntary activity; increased food intake and decreased body temperature appear to play a role in maintaining a higher body weight with increasing age   (MGI Ref ID J:96351)
• obese
  • progressive adult-onset obesity   (MGI Ref ID J:39356)
  • adipose tissue accumulation in the peri-renal, peri-gonadal, and subcutaneous areas of the body   (MGI Ref ID J:39356)
• homeostasis/metabolism phenotype
• *normal* homeostasis/metabolism phenotype
  • blood glucose, insulin, and blood cholesterol levels were within normal ranges   (MGI Ref ID J:39356)
• • decreased circulating follicle stimulating hormone level
    • 4-fold decrease in male mice   (MGI Ref ID J:39356)
  • decreased circulating testosterone level
    • reduced in males observed at 8 to 20 weeks of age   (MGI Ref ID J:39356)
  • decreased core body temperature
    • at >52 weeks of age (but not earlier), female homozygotes exhibit a 1.335 °C reduction in core body temperature relative to wild-type females   (MGI Ref ID J:96351)
    • in contrast, male homozygotes show no significant differences in body temperature at any age relative to wild-type males   (MGI Ref ID J:96351)
• renal/urinary system phenotype
• small male preputial glands
  • preputial glands were either small or absent   (MGI Ref ID J:39356)
• small penis   (MGI Ref ID J:39356)
• reproductive system phenotype
• abnormal Leydig cell morphology
  • reduced number of leydig cells at 32 weeks of age   (MGI Ref ID J:39356)
• abnormal anogenital distance
  • slight reduction in the distance from the penis to the anus   (MGI Ref ID J:39356)
• abnormal estrous cycle
  • 5-month-old female homozygotes display abnormal estrous cycles characterized by a severely truncated estrus and a prolonged diestrus and proestrus   (MGI Ref ID J:101925)
• • prolonged diestrus   (MGI Ref ID J:101925)
  • prolonged proestrus   (MGI Ref ID J:101925)
  • short estrus
    • severely truncated   (MGI Ref ID J:101925)
• abnormal seminal vesicle morphology
  • convoluted   (MGI Ref ID J:39356)
• abnormal spermatogenesis   (MGI Ref ID J:39356)
• • azoospermia
    • absence of spermatozoa in the epididymis at 8 weeks of age   (MGI Ref ID J:39356)
• abnormal vas deferens morphology
  • reduced in size   (MGI Ref ID J:39356)
• absent corpus luteum   (MGI Ref ID J:39356)
• cryptorchism
  • bilateral cryptochism   (MGI Ref ID J:39356)
• decreased litter size
  • females raised under "male free" conditions gave birth to smaller litters   (MGI Ref ID J:39356)
• decreased male germ cell number
  • reduced number or absence of spermatids at 8 weeks of age   (MGI Ref ID J:39356)
• • azoospermia
    • absence of spermatozoa in the epididymis at 8 weeks of age   (MGI Ref ID J:39356)
• decreased ovulation rate
  • in response to exogenous hormones, young female homozygotes ovulate the same number of oocytes as wild-type females   (MGI Ref ID J:101925)
  • in contrast, the number of oocytes released by aged female homozygotes is reduced by >50%, indicating an age-associated reduction in oocytes   (MGI Ref ID J:101925)
  • notably, oocytes from female homozygotes are equally competent for in vitro fertilization assays relative to oocytes from similarly aged wild-type and heterozygous females   (MGI Ref ID J:101925)
• delayed vaginal opening   (MGI Ref ID J:101925)
• early reproductive senescence
  • most mutant females exhibit premature reproductive senescence and support only 3-4 pregnancies during the first few months of a 9-month period   (MGI Ref ID J:101925)
• impaired ovarian folliculogenesis
  • arrested at the early stages of antral formation   (MGI Ref ID J:39356)
• male infertility   (MGI Ref ID J:39356)
• reduced female fertility
  • females raised in the presence of males were fertile, whereas females raised under "male free" conditions had fewer litters   (MGI Ref ID J:39356)
  • in the presence of wild-type or heterozygous males, young female homozygotes (8-12 week-old) can become pregnant and carry litters to full term with no differences in ovulation or litter size relative to age-matched heterozygous and wild-type females   (MGI Ref ID J:101925)
  • however, unlike wild-type females in which fertility averages 8-12 months with ~1 pregnancy per month, mutant females show premature reproductive senescence with most females supporting only 3 to 4 pregnancies during the first few months of a 9-month period   (MGI Ref ID J:101925)
• seminiferous tubule degeneration
  • only remnants of the tubules remained at 32 weeks of age   (MGI Ref ID J:39356)
• small epididymis   (MGI Ref ID J:39356)
• small gonad
  • both male and female homozygotes are hypogonadal with an ~50% reduction or more in gonad size relative to wild-type littermates   (MGI Ref ID J:39356)
• • small ovary
    • females raised in the absence of males exhibited small and pale ovaries   (MGI Ref ID J:39356)
    • size reduction was not apparent until after 4.5 weeks of age   (MGI Ref ID J:39356)
  • small testis
    • significant reduction after 8 weeks of age, though apparent at 4.5 weeks of age   (MGI Ref ID J:39356)
    • testicular tissue was largely replaced by a capsule of fat at 32 weeks of age   (MGI Ref ID J:39356)
• small male preputial glands
  • preputial glands were either small or absent   (MGI Ref ID J:39356)
• small penis   (MGI Ref ID J:39356)
• small seminiferous tubules
  • rudimentary lumens   (MGI Ref ID J:39356)
• small uterus
  • females raised in the absence of males exhibited small uteri with threadlike oviducts   (MGI Ref ID J:39356)
  • size reduction was not apparent until after 4.5 weeks of age   (MGI Ref ID J:39356)
• adipose tissue phenotype
• increased percent body fat
  • at 12-19 weeks (or earlier), male homozygotes show normal percent body fat relative to wild-type males; however, male homozygotes average 11.7% (at 20-51 weeks) and 14.7% (at >52 weeks) more body fat than wild-type males   (MGI Ref ID J:96351)
  • unlike males, female homozygotes exhibit a trend toward increased body fat by 7 weeks of age with significantly increased body fat at 12-19 weeks; female homozygotes average 15.2% (at 20-51 weeks) and 14.9% (at >52 weeks) more fat than wild-type females   (MGI Ref ID J:96351)
• integument phenotype
• small male preputial glands
  • preputial glands were either small or absent   (MGI Ref ID J:39356)

View Research Applications

Research Applications

This mouse can be used to support research in many areas including:

Diabetes and Obesity Research
Obesity Without Diabetes
      adult onset

Endocrine Deficiency Research
Adipose Defects
Gonad Defects
Hypothalamus/Pituitary Defects

Reproductive Biology Research
Developmental Defects Affecting Gonads
Endocrine Deficiencies Affecting Gonads
Fertility Defects

Genes & Alleles

Gene & Allele Information provided by MGI

Allele Symbol		Nhlh2tm1Irk
Allele Name		targeted mutation 1, Ilan R Kirsch
Allele Type		Targeted (knock-out)
Common Name(s)		Nhlh2-;
Mutation Made By		Deborah Good,   Virginia Polytechnic Institute
Strain of Origin		129S4/SvJae
ES Cell Line Name		J1
ES Cell Line Strain		129S4/SvJae
Gene Symbol and Name		Nhlh2, nescient helix loop helix 2
Chromosome		3
Gene Common Name(s)		6230401I09Rik; HEN2; NSCL2; Nscl-2; RIKEN cDNA 6230401I09 gene; bHLHa34;
Molecular Note		The entire coding region was replaced with a PGK-neo cassette. Transcript was undetected by Northern blot analysis of total RNA isolated from homozygous mutant embryos. [MGI Ref ID J:39356]

Genotyping

Genotyping Information

Genotyping Protocols

Nhlh2^tm1Irk,

Separated MCA

Helpful Links

Genotyping resources and troubleshooting

References

References provided by MGI

Selected Reference(s)

Good DJ; Porter FD; Mahon KA; Parlow AF; Westphal H; Kirsch IR. 1997. Hypogonadism and obesity in mice with a targeted deletion of the Nhlh2 gene. Nat Genet 15(4):397-401. [PubMed: 9090387]  [MGI Ref ID J:39356]

Additional References

Nhlh2^tm1Irk related

Cogliati T; Delgado-Romero P; Norwitz ER; Guduric-Fuchs J; Kaiser UB; Wray S; Kirsch IR. 2007. Pubertal Impairment in Nhlh2 Null Mice Is Associated with Hypothalamic and Pituitary Deficiencies. Mol Endocrinol 21(12):3013-3027. [PubMed: 17717072]  [MGI Ref ID J:127370]

Cogliati T; Good DJ; Haigney M; Delgado-Romero P; Eckhaus MA; Koch WJ; Kirsch IR. 2002. Predisposition to arrhythmia and autonomic dysfunction in Nhlh1-deficient mice. Mol Cell Biol 22(14):4977-83. [PubMed: 12077327]  [MGI Ref ID J:78031]

Coyle CA; Jing E; Hosmer T; Powers JB; Wade G; Good DJ. 2002. Reduced voluntary activity precedes adult-onset obesity in Nhlh2 knockout mice. Physiol Behav 77(2-3):387-402. [PubMed: 12419415]  [MGI Ref ID J:96351]

Fox DL; Good DJ. 2008. Nescient helix-loop-helix 2 interacts with signal transducer and activator of transcription 3 to regulate transcription of prohormone convertase 1/3. Mol Endocrinol 22(6):1438-48. [PubMed: 18356286]  [MGI Ref ID J:136071]

Jing E; Nillni EA; Sanchez VC; Stuart RC; Good DJ. 2004. Deletion of the Nhlh2 transcription factor decreases the levels of the anorexigenic peptides alpha melanocyte-stimulating hormone and thyrotropin-releasing hormone and implicates prohormone convertases I and II in obesity. Endocrinology 145(4):1503-13. [PubMed: 14701669]  [MGI Ref ID J:105580]

Johnson SA; Marin-Bivens CL; Miele M; Coyle CA; Fissore R; Good DJ. 2004. The Nhlh2 transcription factor is required for female sexual behavior and reproductive longevity. Horm Behav 46(4):420-7. [PubMed: 15465527]  [MGI Ref ID J:101925]

Wankhade UD; Vella KR; Fox DL; Good DJ. 2010. Deletion of Nhlh2 results in a defective torpor response and reduced Beta adrenergic receptor expression in adipose tissue. PLoS One 5(8):. [PubMed: 20808804]  [MGI Ref ID J:164012]

Health & husbandry

The genotypes of the animals provided may not reflect those discussed in the strain description or the mating scheme utilized by The Jackson Laboratory prior to cryopreservation. Please inquire for possible genotypes for this specific strain.

Health & Colony Maintenance Information

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200.

Colony Maintenance

Breeding & Husbandry	When maintaining a live colony, these mice can be bred as heterozygotes. Homozygous males are infertile. Homozygous females have reduced fertility.

Pricing and Purchasing

Pricing, Supply Level & Notes, Controls

Control Information

	Control
	Wild-type from the colony
	101043 B6129SF1/J	(approximate)
Considerations for Choosing Controls
Control Pricing Information for Genetically Engineered Mutant Strains.

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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX^® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX^® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX^® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

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