ZEB-2021 ·0041-ver9-Araya-jaime_ 1Р

Туре: research-article
ZEBRAFISH
Volume 00, Number 00, 2021 The Fish Haus
© Mary Ann Liebert, lnc.
D01: 10.1089/zeb.2021.0041

Cytogenetics from the Dock:

А New Postmortem Protocol Makes lt PossiЫe
to OЬtain Cytogenetic Preparations from Marine Fish

Cristian Araya-Jaime,1· 2 Claudio Palma-Rojas,2 Natalia Lam,3 and Elisabeth Von Brand 4

Abstract

Cytogenetic studies in marine fish are scarce, and elemental cytogenetic information is availaЫe for not >2% of
the species. Traditional cytogenetic methods require living individuals for their application, making the analysis
of marine ichthyofauna very difficult. In this study, we present а detailed new protocol to obtain cytogenetic
preparations from marine fish, through access to specimens in postmortem condition. The application of this
protocol made it possiЫe to access elemental cytogenetic information (diploid number) in six native species of
the South Pacific Ocean, representative of five orders. In this way, we provide а new low-cost methodological
tool for focused or large-scale cytogenetic analysis, both in economically important, native, or threatened
species.

Keywords: marine fish, cytogenetics protocols, postmortem

с
lntroduction
YTOGENETIC STUDIES IN marine finfish, especially pe-
lagic and benthic, are scarce, and the karyotype of only
marine fish presents many difficulties, due to the fragility of
the specimens, the distance at which they are captured, the
need to have suitaЬle laboratory infrastructure readily avail-
aЫe at the capture site and the time that elapses between
~ 2% of the species is known.1-5 ТЫs lack of elemental Ьi- catch and delivery. 14
ological information gives rise to major knowledge gaps,
In this study, we present а detailed new protocol to obtain
hindering scientific progress in areas such as evolution,
cytogenetic preparations from marine fish through access to
conservation, and natural resource management. Moreover,
specimens in postmortem condition caught Ьу artisanal and
Valente et al. 6 contend that lack of cytogenetic information
sport fishermen. The development of tЫs protocol is based
сап delay the progress of sequencing projects, since these
on the use of а commercial culture medium, РВ-МАХ ТМ
knowledge gaps make it difficult to assemЫe and analyze
(GiЬсо т м ), specially formulated to perform lymphocyte cul-
genomes.
ture in humans and widely used in other vertebrates such as
Traditionally, cytogenetic methods applied to fish have
Ьirds, 15 marine fish, 16 and neotropical canids. 17
used kidney and spleen frajments as target tissues to obtain
metaphase chromosomes.7' In teleost fish, hematopoiesis
Materials and Methods
mainly takes place in the cephalic kidney where stem cells
proliferate to give rise to the various Ыооd ce11 lineages. The То standardize this protocol, eight species were used, re-
spleen also retains the capabllity to originate lymphocytes presenting six orders, purchased mainly from fishermen at the
from lymphoid се\1 precursors.9-12 However, to apply direct port of Coquimbo, СЫ!е (29 ° 57' 18" S 71 ° 20'64" 0 ). The or-
cytogenetic methods, it is first necessary to perform mitotic igin ofthese species was separated into artisanaJ fishing, sport
stimulation followed Ьу the administration of colchicine as fishing, and fish farming. The postmortem time for the spe-
an antirnitotic treatment, all this in live animals. 1•13 Un- cies caught Ьу artisanal fishing was estimated from the time
fortunately, the application of these types of procedures in elapsed since the removal of the nets until the arrival of the

1
Instituto de Investigaciбn Multidisciplinar en Ciencia у Tecnologia, Universidad de La Serena, La Serena, Chile.
2
Laboratorio de Genetica у Citogenetica Vegetal, Departarnento de Biologfa, Universidad de La Serena, La Serena, Chile.
3
Departarnento de Producciбn Animal, FacuJtad de Ciencias Agronбmicas, Universidad de Chile, Santiago, Chile.
4
Departamento de Biologfa Marina Facultad de Ciencias del Mar, Universidad Catбlica del Norte Sede Coquimbo, Coquimbo, Chile.
'ORCID ID (bttps://orcid.org/0000-0003-4639-5207).