Results: 5

Figure 2

Figure 2. From: Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia.

Growth reduction in the Pank2 null animals. Body weight (in grams) of the line 81 homozygous Pank2 mice (open squares) is compared with that of the wild-type (solid squares) littermates as a function of age from weeks 3 to 56. Line graph shows mean ± SEM of body weight. Inset: Representative photograph of a wild-type and Pank2 mouse at week 50. Weight and size differences in the 174 line were similar (data not shown).

Yien-Ming Kuo, et al. Hum Mol Genet. ;14(1):49-57.
Figure 3

Figure 3. From: Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia.

Phenotypic analysis of wild-type and Pank2 mutant testes. (A) Size comparison of representative testis from an 8-week-old wild-type (+/+) male with those from Pank2 mutant (−/−) and heterozygote (+/−) littermates. Identical phenotypes were observed in both the 81 and 174 lines. (B) Histological analysis of testis sections from 8-week-old wild-type (+/+) and mutant (−/−) littermates stained with hematoxylin and eosin (H&E). Images were taken at 20×, 40× and 100× magnification. Arrows indicate mature spermatozoa (40×) and elongated spermatocytes (100×) in the wild-type animals and arrested, round, vaculated spermatids in the mutant. Although the testes of heterozygous mice were of intermediate size, the histochemical analysis of the testis was identical to wild-type.

Yien-Ming Kuo, et al. Hum Mol Genet. ;14(1):49-57.
Figure 5

Figure 5. From: Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia.

Immunohistochemical localization of Pank2 in the testes and retina. (A) Localization of Pank2 in wild-type testes, demonstrating localization to spermatozoa (100×). The cartoon depicts the region of the spermatoza in which mitochondria are concentrated. (B) Localization of Pank2 in the retina (40×). Pank2 is highly expressed in the inner segment (IS) and also in the outer plexiform layer (OPL), inner nuclear layer (INL) and inner plexiform layer (IPL) (right). For reference, a drawing of the rod photoreceptor is presented. No staining is detected in the pre-immune control sera in either tissue (left panel) or in tissues from the mutant animals (middle panel). (C) Localization of Pank1A in the retina, demonstrating staining throughout the photoreceptor, unlike Pank2.

Yien-Ming Kuo, et al. Hum Mol Genet. ;14(1):49-57.
Figure 1

Figure 1. From: Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia.

Generation of Pank2 knock-out animals. (A) Schematic illustration of the targeting strategy. Two portions of the murine Pank2 gene (an 1832 bp fragment containing parts of intron 1 and exon 2 and a 4527 bp fragment spanning exon 3) were incorporated into the targeting vector. Homologous recombination in the regions indicated X resulted in the predicted mutant allele. Correctly targeted ES lines were assessed by PCR using pairs of primers (a+b and c + d). (B) Genotyping of F2 progeny by PCR using two pairs of primers, as described in Materials and Methods. The 446 bp fragment corresponds to the normal gene, and the 344 bp fragment is derived from the correctly targeted gene. Results from wild-type (+/+), heterozygous (+/−) and homozygous knock-out (−/−) mice are shown. (C) RT–PCR demonstrating the presence and absence of the expected 362 bp product from exons 2–4 of brain and testes RNA in wild-type (+/+) and Pank2 (−/−) animals, respectively. White + and − symbols indicate with or without RT prior to PCR. (D) Western blot analysis of proteins from brain and testes extracts of wild-type (+/+) and mutant (−/−) animals with anti-Pank2 antibody. The predicted size for Pank2 is 50 kDa, and a band corresponding to this size is present in wild-type animals but absent in mutant animals.

Yien-Ming Kuo, et al. Hum Mol Genet. ;14(1):49-57.
Figure 4

Figure 4. From: Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia.

Retinal abnormalities in the Pank2 mutant. (A) Representative ERG waveforms recorded from wild-type (+/+) and Pank2 (−/−) littermates at P84 and P411. The scotopic a- and b-wave amplitudes and photopic b-wave amplitude reported in (B) are indicated. Scotopic b-wave amplitudes and photopic b-wave amplitudes are measured in response to a standard flash of +0.4 log cd s/m2, and scotopic a-wave amplitudes are measured in response to a bright flash of +2.4 log cd s/m2 at a 2-fold higher sampling rate, resulting a shorter recorded duration. (B) Summary of ERG amplitude data for all animals tested. Shaded areas delineate wild-type (+/+) mean ± SEM amplitudes for scotopic b-waves (top zone), scotopic a-waves (middle zone) and photopic b-waves (bottom zone). Mean data for the mutant (−/−) are indicated by the open symbols ± SEM The numbers of wild-type and mutant eyes tested were six each at P84, six and 12, respectively, at P285, and 17 and 18, respectively, at P363–P430. Data for all animals of 1 year and older are collapsed into a single time point corresponding to the mean age of P385. (C) Pupillary responses were measured from Pank2 (−/−, open symbol, n = 4) and wild-type (+/+, closed symbol, n = 2) littermate mice, age P285, after dark-adaptation for 90 min and exposed to a 470 nm stimulus. Pupillary area measurements were normalized to the average area measured over a 5 s pre-stimulus baseline and were recorded for 30 s after stimulus presentation. (C, Inset) Pictures showing reduced constriction in Pank2 pupils before and after stimulus when compared with wild-type. (D) Light micrographs from albino wild-type and Pank2 mice at P411. The Pank2 retina shows disorganized and shorter outer segments (OS), shorter inner segments (IS) and a thinner outer nuclear layer (ONL) when compared with age-matched wild-type retina. Area of interest is expanded in insets a and b. Prominent vacuoles are present in the Pank2 OS, indicated by arrows in inset b. (E) Summary of retinal thickness data. Bar graphs show mean ± SEM of the thickness of ONL, IS and OS in wild-type (+/+, solid symbol, n = 15) and Pank2 (−/−, open symbol, n = 19) animals aged P363–P430.

Yien-Ming Kuo, et al. Hum Mol Genet. ;14(1):49-57.

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