Michinari Yokohama

The complete mitochondrial genome of the domestic red deer (Cervus elaphus) of New Zealand and its phylogenic position within the family Cervidae.

We determined the complete nucleotide sequence of the mitochondrial genome of the semidomestic red deer (Cervus elaphus) of New Zealand. The genome was 16,357 bp long and contained 13 protein-coding genes, 12SrRNA, 16SrRNA, 22 tRNAs and a D-loop as found in other mammals. Database homology searches showed that the mitochondrial DNA (mtDNA) sequence from the New Zealand semidomestic deer was similar to partial mtDNA sequences from the European, Norwegian (C. e. atlanticus) and Spanish red deer (C. e. hispanicus). Phylogenetic analysis of the mitochondrial protein-coding regions revealed two well-defined monophyletic clades in subfamilies Cervinae and Muntiacinae. However, red deer and Sika deer were not found to be close relatives. The analysis did identify the red deer as a sister taxon of a Samber/Sika deer clade, although it was more closely related to the Samber than the Sika group.

Genetic relationship among Mongolian native goat populations estimated by blood protein polymorphism

Abstract Several local strains and populations of goats distinguished by morphogenetic and performance characteristics are kept by goat breeders in different natural climatic regions of Mongolia, namely Bayandelger, Ulgii Red, Erchim Black, Dorgon and Zavkhan Buural. The genetic relationships among eight native goat populations in Mongolia at 33 biochemical genetic loci was assessed. A total of 440 animals in eight regional zones were studied. Twelve loci, i.e. the serum transferrin, serum amylase, serum alkaline phosphatase, serum prealbumin-3, cell esterase-D, hemoglobin (Hb) β, hemoglobin (Hb) α-II, cell peptidase-B, cell tetrazolium oxidase, cell esterase-1, cell esterase-2 and cell catalase loci, were found to be polymorphic. The data indicated that Mongolian native goats are not highly differentiated (D=0.0002–0.0038) genetically. To set Mongolian native goats in a larger context, the present data were compared with those on other goat breeds and populations in east and southeast Asia that were previously reported. The average heterozygosity in the Mongolian native goats did not significantly differ from those in other Asian goat populations and breeds. A phylogenetic tree of the gene constitution of the Mongolian native goats and other Asian goat breeds and populations was constructed and revealed that genetically the Mongolian native goats had diverged slightly from the group consisting of Chinese, Japanese, Korean and Indonesian native goats, but markedly from the Indian goat group.

Protein 4.1 expression in the developing hair cells of the mouse inner ear.

Protein 4.1 (band 4.1 or 4.1R) was originally identified as an abundant protein of the human erythrocyte, in which it stabilizes the spectrin/actin cytoskeleton. Subsequently, several new family members, 4.1N, 4.1G and 4.1B, have been identified, which are expressed in many cell types, in particular at cell-cell junctions. We previously reported that 4.1R and 4.1N are expressed in the inner ear hair cells with specific localization patterns, and that 4.1R forms a complex with the membrane-associated guanylate kinase (MAGUK) protein p55 and two deafness gene products, myosin XV and whirlin. To determine the functions of the other family members, 4.1G and 4.1B, we observed their expression patterns in developing stereocilia in mice inner ear hair cells. 4.1G is expressed in the basal tapers of the stereocilia bundle in early postnatal stages. 4.1B was specifically and constantly expressed in the stereocilia tips during postnatal development. Additionally, we found that 4.1B is ablated in the hair cells of both myosin XV and whirlin mutant mice at all stages in hair cell development. These results suggest that 4.1 family members play important roles in the development and maintenance of the inner ear hair cells, and that 4.1B may be a member of the myosin XV-whirlin complex that is important for stereocilia maturation.

Quantitative trait loci on chromosome 5 for susceptibility to frequency-specific effects on hearing in DBA/2J mice

The DBA/2J strain is a model for early-onset, progressive hearing loss in humans, as confirmed in the present study. DBA/2J mice showed progression of hearing loss to low-frequency sounds from ultrasonic-frequency sounds and profound hearing loss at all frequencies before 7 months of age. It is known that the early-onset hearing loss of DBA/2J mice is caused by affects in the ahl (Cdh23ahl) and ahl8 (Fscn2ahl8) alleles of the cadherin 23 and fascin 2 genes, respectively. Although the strong contributions of the Fscn2ahl8 allele were detected in hearing loss at 8- and 16-kHz stimuli with LOD scores of 5.02 at 8 kHz and 8.84 at 16 kHz, hearing loss effects were also demonstrated for three new quantitative trait loci (QTLs) for the intervals of 50.3–54.5, 64.6–119.9, and 119.9–137.0 Mb, respectively, on chromosome 5, with significant LOD scores of 2.80–3.91 for specific high-frequency hearing loss at 16 kHz by quantitative trait loci linkage mapping using a (DBA/2J × C57BL/6J) F1 × DBA/2J backcross mice. Moreover, we showed that the contribution of Fscn2ahl8 to early-onset hearing loss with 32-kHz stimuli is extremely low and raised the possibility of effects from the Cdh23ahl allele and another dominant quantitative trait locus (loci) for hearing loss at this ultrasonic frequency. Therefore, our results suggested that frequency-specific QTLs control early-onset hearing loss in DBA/2J mice.

Development and characterization of 260 microsatellite loci in the domestic goat, Capra hircus.

Comments: The current study emphasized genes with lipogenic activity and their relationship to reproductive events. Weaning-to-oestrus interval was associated with five SNPs. A single THRSP SNP maintained an association with WEI after correction testing. THRSP has been identified as a murine candidate gene within overlapping QTL for the effects of lifetime fertility and dietary restriction. Data generated from this study imply that SNPs within lipogenic genes could be used to assist with identifying animals that tolerate the metabolic demands of lactation while maintaining reproductive competence.

Expression analysis of the type I keratin protein keratin 33A in goat coat hair

The coat of a goat, like that of many mammalian species, consists of an outer coat of coarse hairs and an under coat of fine, downy hairs. The coarse guard hairs are produced by primary follicles and the finer cashmere hairs by secondary follicles. We previously reported that hair keratins are components of cashmere hair, and proteomic analysis revealed that their expression is elevated in winter coat hair. To determine detailed characterization, we have cloned keratin 33A gene, a major highly expressed keratin in winter, and then analyzed the expression of goat hair coat. By Western analysis, we detected that keratin 33A protein is expressed only in hair coat among the various goat tissues. Moreover, the expression level in winter has increased in cashmere high-producing Korean native breed, whereas the expression levels between summer and winter had not changed in cashmere low-producing Saanen. In addition, by immunohistochemistry we determined that keratin 33A is localized in the major cortex portion of cashmere fiber. These results confirm that keratin 33A is a structural protein of goat cashmere hair fiber.

Subgrouping of equine transferrin types by peptide mapping

quantitative variants between T+ and Tcomponents composed of Tf types, the Tf types could be classified into three groups as Tf-D・F・H, Tf-O・R and Tf-X subgroups. To certify furthermore the results, the equine Tf types were subgrouped from peptide cleavage patterns produced by peptide mapping using four proteases (α-chymotrysin, V8 protease, papain and endoprotease Asp-N). The results obtained are as follows: 1) When Tf・D, F and H types were treated by the four proteases, these three Tf types had almost the same peptide cleavage patterns. Although there were respectively some different band patterns between each peptide cleaved in Tf・O and R types, they had more mutually similar points than those of the Tf-D・F・H subgroup. This means that Tf・O and R types may be mutually specialized, but it was guessed that they can be basically classified into the same subgroup in view of having many of the same peptide cleavage components. The four peptide patterns which were cleaved donkeys Tf components, were each very similar with those of equine Tf・O and R types. 2) T+ and Tcomponents of Tf・D and R types were each cleaved by V8 protease and α-chymotrypsin, and both components of the Tf・D type showed almost same peptide pattern, and there were distinct differences when the two components of the Tf・R type were digested by the latter protease. From analysis by peptide mapping, Tf・O and R types might be newly subgrouped, but it was again recognized that the peptide structures of the Tf・O and R types were each distinctly different from those of the Tf-D・F・H subgroup. Accordingly, the equine Tf types could be classified into two groups as Tf-D・F・H and Tf-O・R types by peptide mapping. Then the T+ and Tcomponents in the Tf・R type had basically different peptide structure, respectively.

Developmental Changes of Transferrin Concentration in Light-breed Horses

軽種馬のトランスフェリン(Tf)は,生直後と生後1ヵ月齢前後に鉄飽和またはそれに極めて近い状態になる時があることが二次元電気泳動(2-D)法によるTfのパターン分析で分った.また,この時期はTf量および血清鉄値もピークを示し,Tfの総鉄結合能が高くなっていた.一方免疫グロブリン量とTfの加齢変動を比較した結果,IgG量が最も低下する生後1～3ヵ月齢頃は,Tf量が成体時の約2倍量保持している時期であった.以上のことから,Tfの加齢変動は,生体内におけるTfの鉄運搬機能および感染防御と関連した成育時期におけるTfの顕著な作用増進の現象と考えられた.

On the Hemoglobin Types of Japanese Native Horses

デン粉ゲル電気泳動法,6M尿素加デン粉ゲル電気泳動法および等電点電気泳動法(IEF)を用い,検出された各Hb型間の関係を分析するとともに,日本在来馬種におけるHb型の遺伝子ならびにhaplotypeの頻度を調査した結果,次のごとき成績が得られた.1) BRAEND and STORMONTの分類法によるHbAおよびHbA遺伝子はそれぞれHb-α1(dup.)およびHb-α0(dup.)遺伝子と一致した.また,Hba(またはHb-α0)遺伝子をもつ個体には特異的にIEF法によるhaplotype Aが認められた.2) Hba(またはHb-α0)遺伝子およびhaplotype Aは日本在来馬種のうち,北海道和種馬,木曽馬,トカラ馬および与那国馬に認められ,トカラ馬における頻度が最も高く0.533であった.3) IEF法によって検出されるHbのhaplotypeでは,野間馬はBI/BI型のみであった.トカラ馬にはhaplotype BIは認められず,BIIとAでそれぞれ46.7%および53.3%であった.また,御崎馬にはhaplotype Aは認められず,BIとBIIがそれぞれ43.8%および56.3%であった.

Variation of the Amount of Serum Transferrin Protein in Horses

馬血清トランスフェリン(Tf)蛋白量の差異から,通常のTf型とsilent geneをもつTf型をより明確に識別するため,各要因によるTf蛋白量の変動について調べた.すなわち,検体の採取時の違いによる季節的並びに個体内の年間の変動について調査すると同時に,Tf型の違いおよび品種の達いによって異なるかどうかについて調べた.その成績は次の如くである.1, 通常のTf型の個体内におけるTf蛋白量変動の最高値と最低値との差(変動幅)は約32～92mg/dlの間,また,最高値および最低値と平均値間の差はそれぞれ14.8～53.7mg/dlの間であった.一方,silent geneをもつTf型ではその変動幅は小さく約25～32mg/dlの間,また,後者の平均値との間における差は8.9～18mg/dlの間であった.2, 調査に用いた12個体のTf蛋白量の測定値はいずれもP=0.05における棄却限界の範囲内であり,それらの変異係数は3.7～8.7%であった.3, 季節的にはTf蛋白量に著しい変動が認められなかった.4, 軽種馬における15型のTf型間のTf蛋白量は,易動度の速いD,FおよびHバンドの組合わせからなるDF型,FF型,FH型およびHH型が平均値で約370～390mg/dlの間にあり,高い値を示すグループであったのに対し,易動度の遅いRバンドを含むDR型,FR型,OR型およびRR型が平均値で約310～330mg/dlの間にあり低い値を示すグループであった.これら2群の間には有意差が認められた(P<0.001)。5, ホモ型のTf蛋白量はFF型,HH型,OO型,DD型およびRR型の順に高く,それぞれの平均値は382.4±2.9, 370.1±9.7, 352.6±5.2, 347.7±1.2および312.3±7.9mg/dlであった,6, Tf型間にTf蛋白量の差が存在することから,silent geneをもつTf型を識別する場合は通常のホモ型と,これに対応するsilent geneをもつTf型との間で,Tf蛋白量を比較する方がより明確な識別が期待できる.7, サラブレッド種,アングロアラブ種,重種,北海道和種,木曽馬およびポニー種の6品種間には明確なTf蛋白量の差は認められなかった.