Kat is being used extensively in many countries as a central nervous system stimulant. The effect of three doses of crude kat extract on chromosomal division and abnormalities in bone marrow, as well as on DNA, RNA, and total protein content in brain and liver was studied in laboratory rats in order to test the possible mutagenicity of the drug. Kat was given as a single subcutaneous injection at 0.05 (usage dose), 0.52 (intermediate dose), and 1.00 (sublethal dose) g/kg body weight. Animals were sacrificed at 6, 24, and 48 hr after treatment. Also, some animals were exposed subacutely for 5 consecutive days with sacrifice occurring 6 hr after the last injection. The mitotic index was reduced by all treatments, with the greatest effect occurring in the subacute treatment. Chromosomal abnormalities were induced by kat at all three doses, administered acutely or subacutely. The significant chromosomal aberrations were in the form of gaps, breaks, centromeric attenuations, and centric fusions. The concentration of DNA, RNA, and total protein in liver and brain decreased at all doses, with the greatest decrease occurring after subacute treatment. These findings suggest that kat has a profound effect on cell proliferation, on chromosomal abnormalities, and on DNA, RNA, and total protein synthesis.

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Ninety-nine loci have been assigned to river buffalo chromosomes, 67 of which are coding genes and 32 of which are anonymous DNA segments (microsatellites). Sixty-seven assignments were based on cosegregation of cellular markers in somatic cell hybrids (synteny), whereas 39 were based on in situ hybridization of fixed metaphase chromosomes with labeled DNA probes. Seven loci were assigned by both methods. Of the 67 assignments in somatic cell hybrids, 38 were based on polymerase chain reaction (PCR), 11 on isozyme electrophoresis, 10 on restriction endonuclease digestion of DNA, 4 on immunofluorescence, and 4 on chromosomal identification. A genetic marker or syntenic group has been assigned to each arm of the five submetacentric buffalo chromosomes as well as to the 19 acrocentric autosomes, and the X and Y chromosomes. These same markers map to the 29 cattle autosomes and the X and Y chromosomes, and without exception, cattle markers map to the buffalo chromosome or chromosomal region predicted from chromosome banding similarity.

To identify the river buffalo chromosome carrying the genes coding for GAPD, TPI1, and LDHB, karyotypic examination was carried out on 14 buffalo-hamster hybrid clones previously tested for presence of this syntenic group. In cattle, this group (U3) has been assigned to chromosome 5, which is assumed to be homologous to the long arm of buffalo chromosome 4. Chromosome 4 was present in all five clones expressing the three enzymes, and absent in all seven negative clones, indicating that in the buffalo GAPD, TPI1, and LDHB are located on chromosome 4. One clone, expressing GAPD and TPI1, but not LDHB, was found to carry a translocation between hamster marker chromosome M(2) and buffalo 4q1 → 4qter. In another clone, expressing LDHB, but not GAPD and TPI1, chromosome 4 was absent, while a very small, unidentifiable acrocentric was present. These observations suggest that LDHB is located in the proximal part of 4q1, and that GAPD and TPI1 are located more distally, in 4q1 → 4q2. ZUSAMMENFASSUNG: Lokalisierung von Genen auf Chromosom 4 des Flußbüffels durch Büffel-Hamster-Hybridzellen Zur Identifikation von Flußbüffelchromosomen mit Genen für GAPD, TPI1 und LDHB wurden Karyotypenbestimmungen an 14 Büffel-Hamster-Hybridklonen durchgeführt, die vorher auf Anwesenheit der betreffenden synthenischen Gruppen geprüft worden waren. Bei Rindern wird diese Gruppe (U3) dem Chromosom 5 zugeordnet, welches als homolog mit dem langen Arm des Büffelchromosoms 4 betrachtet wird. Chromosom 4 war in allen fünf Klonen, die die drei Enzyme exprimiert haben, vorhanden und fehlte in allen sieben negativen klonen, so daß angenommen werden kann, daß sich bei Büffeln GAPD, TPI1 und LDHB auf Chromosom 4 befinden. Bei einem Klon, der GAPD und TPI1, aber nicht LDHB zeigte, wurde eine Translokation zwischen dem Hamstermarkerchromosom M2 und Büffel 4q1 → 4qter gefunden. Im einem anderen Klon, der LDHB, nicht aber GAPD und TPI1 zeigte, war Chromosom 4 nicht vorhanden, wohl aber ein sehr kleines, nicht identifizierbares akrozen-trisches Chromosom. Diese Beobachtungen weisen darauf hin, daß sich LDHB im proximalen Teil von 4q1 befindet und GAPD und TPI1 vertu distal in 4q1 → 4q2 lokalisiert sind.

The cosegregation of ten coding loci has been investigated, in a panel of 37 somatic cell hybrids resulting from the fusion of a hamster cell line and river buffalo lymphocytes, by use of Southern hybridization technique. Five syntenic groups, TCRB-PGY3, ASS-ABL, FUCA1P-CRYG, MBP-YES1, and CGN1-ACTA1, previously assigned to cattle as U13, U16, U17, U28, and U29 respectively, were also found to be syntenic in buffalo. Based on the extensive syntenic conservation and banding homology between cattle and river buffalo, comparative mapping predicts the localization of these syntenic groups on river buffalo Chromosomes (Chrs) :BBU7, BBU12, BBU2q, BBU22, and BBU4q respectively as they have been previously localized on cattle Chrs BTA4, BTA11, BTA2, BTA24 & BTA28.

Forty-five somatic buffalo-hamster hybrid clones were analysed for the presence or absence of PCR products of 10 primers. Five syntenic groups were identified: CGA-D9S1; CD18-D1S4; OXT-PRNP; LDLR-D7S3; BSPN-D14S2. These same syntenic groups were reported to be syntenic in cattle and were assigned to U2 (BTA 9), U10 (BTA 1), U11 (BTA 13), U22 (BTA 7), and U24 (BTA 14), respectively. Based on chromosomal homology between cattle and river buffalo chromosomes, these syntenic groups are expected to be assigned to buffalo chromosomes BBU 10, BBU 1q, BBU 14, BBU 9, and BBU 15, respectively. ZUSAMMENFASSUNG: Genkartierung beim Flußbüffel (Bubalis bubalis L.): Fünf Syntäniegruppen Fünfundvierzig somatische Büffel-Hamster Hygrid Klone wurden bezüglich An-oder Abwesenheit von PCR Produkten von 10 primern analysiert. Es wurden 5 Syntäniegruppen identifiziert: CGA-D9S1; CD18-D1S4; OXT-PRNP; LDLR-D7S3; BSPN-D14S2. Dieselben Syntäniegruppen wurden in Rindern gfunden und zugeordnet zu U2(BTA 9), U10(BTA 1), U11(BTA 13), U22(BTA 7) und U24(BTA 14). Auf grund chromosomaler Homolgie zwischen beiden Arten sollten die Syntäniegruppen auf den Büffelchromosomen BBU 10, BBU 1q, BBU 14, BBU 9, und BBU 15 liegen.

In the process of developing a buffalo physical map using somatic cell hybrids and the cattle gene map as a template, ten PCR primers designed for four coding genes: F10, FSHB, HBB, and CYM and six DNA segments: TGLA9, TGLA227, UWCA5, CSSM6, CSSM47 and RF131 were tested on a panel of 47 buffalo-hamster somatic cell hybrids. F10-TGLA9, FSHB-HBB and UWCA5-TGLA227, respectively, were found to segregate together forming three syntenic groups. These three syntenic groups have also been reported in cattle, where they have been assigned to chromosomes BTA 12, BTA 15 and BTA 18, respectively. Comparative mapping predicts the assignment of these syntenic groups to river buffalo chromosomes BBU 13, BBU 16 and BBU 18, respectively. © Inra/Elsevier, Paris buffalo / chromosome / synteny / PCR markers / gene mapping * Correspondence and reprints Résumé-Assignation de marqueurs PCR aux chromosomes du buffle de rivière. Dans le but de développer la carte génétique physique du buffle en utilisant l'hybridation cellulaire somatique et la carte génétique bovine comme référence, dix amorces PCR correspondant à quatre gènes codants : F10, FSHB, HBB et CYM et six segments d'ADN : TGLA9, TGLA227, UWCA5, CSSM6, CSSM47 et RF131 on été testés sur une série de 47 hybrides cellulaires somatiques entre buffle et hamster. F10-TGLA9, FSHB-HBB et UWCA5-TGLA227, respectivement, ont été trouvés ségréger ensemble pour former trois groupes synténiques. Ceux-ci ont été aussi observés chez les bovins où ils ont été assignés aux chromosomes BTA 12, BTA 15 et BTA 18, respectivement. Les cartes comparées permettent de prédire une assignation respective des trois groupes synténiques aux chromosomes BBU 13, BBU 16 et BBU 18 du buffle de rivière. @ Inra/Elsevier, Paris bufHe / chromosome / synténie / marqueurs PCR / carte génétique

A standard karyotype for the River Buffalo has recently been established. The largest five chromosomes are biarmed and, based on the banding homology between cattle and buffalo chromosomes, were suggested to originate from the fusion of cattle acrocentric chromosomes. The origin of buffalo chromosome 1 is controversial due to the difficulty in differentiating between the small acrocentric cattle chromosomes. Using molecular markers assigned to cattle chromosomes, synteny between CD18, a marker for BTA1, and markers for small acrocentric cattle chromosomes BTA 24 to BTA 29 was investigated in buffalo/hamster somatic cell hybrids. The investigation revealed that CD18 is syntenic with ANT1, a marker for cattle chromosome 27. The present results confirm that buffalo BBU1 results from fusion of cattle BTA 1 and BTA 27. They also underline the importance of biarmed buffalo chromosomes for the identification of small cattle acrocentrics.