Iranian Journal of Microbiology 2013. 5(1):91-98.

A new approach for aggregation of Paramecium caudatum by nitric oxide.
Manizheh Karami, Seyed Sajad Shahrokhi, Bahram Kazemi, Seyedeh Samaneh Moezzi


Background and Objective: Nitric oxide (NO) plays a role in thermoregulation and growth of protozoa. This work aimed to add the molecule NO in physiology of protozoa in contact with abused narcotic substances.
Materials and Methods: A sedative drug, morphine, was infused into a cell chamber containing Paramecia. The cell response to the drug was recorded promptly after drug infusion using a potency protocol provided for the first time at this laboratory. A precursor of NO, L-arginine, was treated jointly with drug to involve the NO system in protozoan performance to drug exposure. Marking of NADPH-diaphorase (NADPH-d) was followed to provide data to explain the mechanisms.
 Results: Morphine, particularly 0.5 to 60 μg/μl, aggregated the Paramecia. The infusion of L-arginine (1 to 8 µg/µl) together with morphine potentiated this effect, though, pre-usage of L-NAME (1 to 8 µg/µl), a blocker of NO production, reversed the response. Notably the activation of NADPH-d in solely morphine or L-arginine plus morphine samples was revealed. However, the expression of marker was attenuated upon pre-infusion with L-NAME.
Conclusion: This study introduces a new approach to involve NO in physiology of aggregation of Paramecia following exposure to the misused sedative drug, morphine.


Aggregation; Morphine; NADPH-diaphorase; Nitric oxide; Paramecium

Full Text:



Berger AC, Whistler JL. How to design an opioid drug that causes reduced tolerance and dependence. Ann Neurol 2010; 67: 559-569.

Klein G, Rossi GC, Waxman AR, Arout C, Juni A, Inturrisi CE, et al. The contribution of MOR-1 exons 1-4 to morphine and heroin analgesia and dependence.Neurosci Lett 2009; 457: 115-119.

Jardinaud F, Roques BP, Noble F. Tolerance to the reinforcing effects of morphine in delta9- tetrahydrocannabinol treated mice. Behav Brain Res 2006; 173: 255-261.

Lanteri C, Salomon L, Torrens Y, Glowinski J, Tassin JP. Drugs of abuse specifically sensitize noradrenergic and serotonergic neurons via a non- dopaminergic mechanism. Neuropsychopharmacol 2008;33: 1724-1734.

Garthwaite J, Glutamate, nitric oxide and cell –cell signaling in the nervous system. Trends Neurosci 1991;14: 60-67.

Moncada S, Palmer RMJ, Higgs EA. Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 1991; 43: 109-142.

Yahyavi-Firouz-Abadi N, Tahsili-Fahadan P, Ghahremani MH, Dehpour AR. Melatonin enhances the rewarding properties of morphine: involvement of the nitric oxidergic pathway. J Pineal Res 2007; 42: 323-9.

Zarrindast MR, Askari E, Khalilzadeh A, Nouraei N.Morphine state-dependent learning sensitization and interaction with nitric oxide. Pharmacology 2006; 78:66-71.

Fukuto JM, Mayer B (1996). The enzymology of nitric oxide synthase.In: Feelisch M, Stamler JS, Editors. Methods in Nitric Oxide Research. Wiley & Sons. New York.

Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J 1992; 6: 3051-3064.

Nighorn A, Gibson NJ, Rivers DM, Hildebrand JG, Morton DB. The nitric oxide-cGMP pathway may mediate communication between sensory afferents and projection neurons in the antennal lobe of Manduca sexta. J Neurosci 1998; 18: 7244-7255.

Goldstein J, Paveto C, Lopez-Costa JJ, Pereira C, Alonso G, Torres HN, Flawia MM. Immuno and cytochemical localization of Trypansoma cruzi nitric oxide synthase. Biocell 2000; 24: 217-222.

Piacenza L, Peluffo G, Radi R. L-arginine-dependent suppression of apoptosis in Trypanosoma cruzi: contribution of the nitric oxide and polyamine pathways. Proc Natl Acad Sci USA 2001; 98: 7301-7306.

Malvin GM, Cecava N, Nelin LD. Nitric Oxide Production and Thermoregulation in Paramecium caudatum. Acta Protozool 2003; 42: 259-267.

Cadet P, Stefano GB.Mytilus edulis pedal ganglia express µ opiate receptor transcripts exhibiting high sequence identity with human neuronal m1. Mol Brain Res 1999; 74: 242.

Kream RM, Zukin RS, Stefano GB. Demonstration of two classes of opiate binding sites in the nervous tissue of the marine mollusc Mytilus edulis: positive homotropic cooperativity of lower affinity binding sites. J Biol Chem 1980; 255: 9218.

Stefano G B, Scharrer B. High affinity binding of an enkephalin analog in the cerebral ganglion of the insect Leucophaea maderae (Blattaria). Brain Res 1981; 225: 107.

Liu Y, Shenouda D, Bilfinger TV, Stefano M L, Magazine H I, Stefano G B. Morphine stimulates nitric oxide release from invertebrate microglia. Brain Res.1996; 722: 125.

Stefano GB, Scharrer B. The presence of the m3 opiate receptor in invertebrate neural tissues. Comp Biochem Physiol 1996; 113C: 369.

Thomas E, Pearse AG. Solitary active cells. Histochemical demonstration of damage-resistant nerve cells with a TPN-diaphorase reaction. Acta Neuropathol 1964; 3: 238-49.

Karami M, Baloutch M. A systematic study on freshwater ciliates in Tehran region. Daneshvar Med J1999; 6: 29-32 (In Persian).

Karami M. The biosystematic of freshwater protozoa (1992). The MSc Thesis at the University of Tehran. Tehran. Iran.

Finlay BJ, Rogerson A, Cowling AJ (1988). A beginner's guide to the collection, isolation, and cultivation of freshwater protozoa. Freshwater Biological Association. Ambleside. Cumbria. UK.

Karami M. Dichotomous key for Paramecium spp. & Vorticella spp in freshwaters of Tehran region.Daneshvar Med J 2001; 8: 135-138.

Laybourn-Parry J (1984). A functional biology of free- living protozoa. Biddles Ltd. Guildford and King's Lynn. London. UK.

Weinberg RJ, Valtschanoff JG. Schmidt HHHW (1996). The NADPH diaphorase histochemical staining. In: Ferelisch M, Stamler J, Editors. Methods in Nitric Oxide Research. John Wiley & Sons Ltd. New York, p. 237-248.

Andronowska A, Chruściel M. Expression and cellular distribution of NADPH-diaphorase and nitric oxide synthases in the porcine uterus during early pregnancy. Folia Histochem Cytobiol 2007; 45: 375-80.

Imada C, Oosawa Y. Thermoreception of Paramecium: different Ca2+ channels were activated by heating and cooling. J Mem Biol 1999; 168: 283-287.

Linder JU, Engel P, Reimer A, Kruger T, Plattner H, Schultz A, et al. Guanylyl cyclases with the topology of mammalian adenylyl cyclases and an N-terminal P-type ATPaselike domain in Paramecium, Tetrahymena and Plasmodium. EMBO J 1999; 18: 4222-4232.

Prajer M, Fleury A, Laurent M. Dynamics of calcium regulation in Paramecium and possible morphogenetic implication. J Cell Sci 1997; 110: 529-535.

Preston RR, Saimi Y, Kung C. Calcium current activated upon hyperpolarization of Paramecium tetraurelia. J Gen Physiol 1992; 100: 233-251.

Thiele J, Schultz JE. Ciliary membrane vesicles of Paramecium contain the voltage-sensitive calcium channel. Proc Nat Acad Sci USA 1981;78: 3688-3691.

Marino JM, Sherman GT, Wood DC. Partial Cloning of Putative G-Proteins Modulating Mechanotransduction in the Ciliate Stentor. J Eukaryotic Microbiol 2001; 48:527-536.

Colasanti M, Venturini G. Nitric Oxide in Invertebrates.Mol Neurobiol 1998; 7: 157-174.

Amaroli A, Ognibene M, Trielli F, Trombino S, Falugi C, Corrado_ Umberta Delmonte M. Detection of NADPH- diaphorase activity in Paramecium primaurelia. Eur. J Protistol 2006; 42: 201-208.

Zarrindast MR, Karami M, Sepehri H, Sahraei H. Influence of nitric oxide on morphine-induced conditioned place preference in the rat central amygdale. Eur J Pharmacol 2002; 453: 81-89.

Zarrindast MR, Karami M, Sepehri H, Sahraei H. Role of nitric oxide in the rat hippocampal CA1 area on morphine-induced conditioned place preference. Eur J Pharmacol 2002; 449: 113-119.

Hashemi M, Karami M, Zarrindast MR, Sahebgharani M. Role of nitric oxide in the rat hippocampal CA1 in morphine antinociception. Brain Res 2010; 1313: 79-88.


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.