Extravisual opsins in the blind shrimp <i>Creaseria morleyi</i>: presence and expression

The presence of a long-wavelength sensitive (LWS) opsin gene was demonstrated in the stygobitic crustacean Creaseria morleyi (Creaser, 1936) by PCR readings from genomic DNA. In order to find the expression of this gene in extraocular tissue, shrimps were collected and placed in a tank to expose them to light/dark conditions for a period of 72 hours, and immediately after, sacrificed and sampled for RNA in the eyes, cephalothorax, abdomen, and sixth abdominal segment plus uropods. The transcripts of the LWS opsin gene were found in the eyes and abdomen of individuals exposed to light. The expression of these opsins could be involved in extravisual functions such as synchronization of their biological processes with environmental cycles related to diurnal vertical migration.

Keywords:
Photopigment; phototransduction; stygofauna

Authorship

José R. Pérez-Calderón

Performed research

Analysis and interpretation of data

Preparation of figures

Writing - original draft

Maestría en Ciencias Orientación Genómica. Departamento de Ciencias Químico-Biológicas, Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez. Ciudad Juárez, México. Av. Plutarco Elías Calles 1210, 32310.Universidad Autónoma de Ciudad JuárezMéxicoCiudad Juárez, México Maestría en Ciencias Orientación Genómica. Departamento de Ciencias Químico-Biológicas, Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez. Ciudad Juárez, México. Av. Plutarco Elías Calles 1210, 32310.

https://orcid.org/0000-0002-0767-8995

Jorge A. Pérez-León

Conceptualization and Design

Acquisition of data

Analysis and interpretation of data

Writing - critical review & editing

https://orcid.org/0000-0002-9148-2054

Nuno Simões

Conceptualization and Design

Performed research

Acquisition of data

Writing - critical review & editing

Unidad Multidisciplinaria de Docencia e Investigación UMDI-Sisal, Facultad de Ciencias, Universidad Nacional Autónoma de México. Sisal, Yucatán, México. 97356.Universidad Nacional Autónoma de MéxicoMéxicoSisal, Yucatán, MéxicoUnidad Multidisciplinaria de Docencia e Investigación UMDI-Sisal, Facultad de Ciencias, Universidad Nacional Autónoma de México. Sisal, Yucatán, México. 97356.

Laboratorio de Resiliencia Costera (LANRESC, CONACYT). Sisal, Yucatán, MéxicoLaboratorio de Resiliencia CosteraMéxicoSisal, Yucatán, MéxicoLaboratorio de Resiliencia Costera (LANRESC, CONACYT). Sisal, Yucatán, México

International Chair for Coastal and Marine Studies in Mexico, Harte Research Institute for Gulf of Mexico Studies, Texas A&M University-Corpus Christi, TX. United States of AmericaTexas A&M UniversityUnited States of AmericaCorpus Christi, TX, United States of AmericaInternational Chair for Coastal and Marine Studies in Mexico, Harte Research Institute for Gulf of Mexico Studies, Texas A&M University-Corpus Christi, TX. United States of America

https://orcid.org/0000-0001-7490-3147

Marisela Aguirre-Ramírez

Conceptualization and Design

Analysis and interpretation of data

Preparation of figures

Writing - critical review & editing

Cuerpo Académico Consolidado Biología Celular y Molecular, Departamento de Ciencias Químico-Biológicas, Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez. Ciudade Juárez, México. Av. Plutarco Elías Calles 1210, 32310.Universidad Autónoma de Ciudad JuárezMéxicoCiudade Juárez, MéxicoCuerpo Académico Consolidado Biología Celular y Molecular, Departamento de Ciencias Químico-Biológicas, Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez. Ciudade Juárez, México. Av. Plutarco Elías Calles 1210, 32310.

https://orcid.org/0000-0002-9764-0324

Roxana E. Malpica-Calderón

Conceptualization and Design

Analysis and interpretation of data

Preparation of figures

Writing - critical review & editing

https://orcid.org/0000-0002-4933-3071

Alejandro Botello

Conceptualization and Design

Analysis and interpretation of data

Writing - critical review & editing

https://orcid.org/0000-0003-1915-2920

CORRESPONDING AUTHOR
Alejandro Botello alejandro.botello@uacj.mx

JRPC
E-mail: jrpc.64@gmail.com

JAPL
E-mail: alberto.perez@uacj.mx

MAR E-mail: marisela.aguirre@uacj.mx

REMC
E-mail: roxana.malpica@uacj.mx

AB E-mail: alejandro.botello@uacj.mx

NS E-mail: ns@ciencias.unam.mx

Associate Editor:

Christopher Tudge

Competing interests

All authors declare that they have no conflicts of interest.

SCIMAGO INSTITUTIONS RANKINGS

Figures

Figures (3)

Thumbnail

Figure 1.
Opsins cluster diagram. UPGMA clustering diagram with bootstrap values of 1000 replicates for each node. The lower bar indicates the genetic distance. The Creaseria morleyi opsin is in the red square, in the group of the long wavelength sensitive opsins of crustaceans. LW: long-wavelength sensitive opsin; MW: medium-wavelength sensitive opsin; SW: short-wavelength sensitive opsin; opn: opsin.

Thumbnail

Figure 2.
Amino acid sequence alignment of the Creaseria morleyi long wavelength opsin and other crustacean opsins. The conserved residues are shadowed, and the gaps indicated by dash (-). Amino acid numbers per individual sequence are indicated on the right. The transmembrane helix is shown as roman numeral and enclosed in squares. The “DRY” and “QAKKM” motif, which are important for the union to the G protein are shown by # and % symbols respectively. The cysteine residues responsible of the disulfide bond stabilization are indicated by ! symbol. The glutamic acid that acts as a counterion on the Schiff base is shown as $. The Y and S amino acids with the LWS/MWS spectral sensitivity are shown with the symbol &.

Thumbnail

Figure 3.
Electrophoresis of cDNA of amplified long wavelength opsin products by PCR from cDNA obtained by RT-PCR performed on RNA samples of Creaseria morleyi. cDNA from the sixth abdominal segment plus uropods (6A+U), abdomen (A), eyes (E), and cephalothorax (C), showed PCR products using specific primers for long wavelength opsin 1. The smallest band shown from the DNA ladder (L) is 500 bp, a 100 bp distance separates each band between the band corresponding to 500 bp ant and the band of 1000 bp. The expected amplicon was of 848 pb and can be observed in eyes and abdomen in continuous light condition, and in abdomen and in sixth abdominal segment plus uropods in total darkness condition.

Figure 1. Opsins cluster diagram. UPGMA clustering diagram with bootstrap values of 1000 replicates for each node. The lower bar indicates the genetic distance. The Creaseria morleyi opsin is in the red square, in the group of the long wavelength sensitive opsins of crustaceans. LW: long-wavelength sensitive opsin; MW: medium-wavelength sensitive opsin; SW: short-wavelength sensitive opsin; opn: opsin.

Figure 2. Amino acid sequence alignment of the Creaseria morleyi long wavelength opsin and other crustacean opsins. The conserved residues are shadowed, and the gaps indicated by dash (-). Amino acid numbers per individual sequence are indicated on the right. The transmembrane helix is shown as roman numeral and enclosed in squares. The “DRY” and “QAKKM” motif, which are important for the union to the G protein are shown by # and % symbols respectively. The cysteine residues responsible of the disulfide bond stabilization are indicated by ! symbol. The glutamic acid that acts as a counterion on the Schiff base is shown as $. The Y and S amino acids with the LWS/MWS spectral sensitivity are shown with the symbol &.

Figure 3. Electrophoresis of cDNA of amplified long wavelength opsin products by PCR from cDNA obtained by RT-PCR performed on RNA samples of Creaseria morleyi. cDNA from the sixth abdominal segment plus uropods (6A+U), abdomen (A), eyes (E), and cephalothorax (C), showed PCR products using specific primers for long wavelength opsin 1. The smallest band shown from the DNA ladder (L) is 500 bp, a 100 bp distance separates each band between the band corresponding to 500 bp ant and the band of 1000 bp. The expected amplicon was of 848 pb and can be observed in eyes and abdomen in continuous light condition, and in abdomen and in sixth abdominal segment plus uropods in total darkness condition.

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Extravisual opsins in the blind shrimp <i>Creaseria morleyi</i>: presence and expression

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[1] CORRESPONDING AUTHOR
Alejandro Botello alejandro.botello@uacj.mx